Genetic studies on the role of type IA DNA topoisomerases in DNA metabolism and genome maintenance in Escherichia coli

Le surenroulement de l’ADN est important pour tous les processus cellulaires qui requièrent la séparation des brins de l’ADN. Il est régulé par l’activité enzymatique des topoisomérases. La gyrase (gyrA et gyrB) utilise l’ATP pour introduire des supertours négatifs dans l’ADN, alors que la topoisomérase I (topA) et la topoisomérase IV (parC et parE) les éliminent. Les cellules déficientes pour la topoisomérase I sont viables si elles ont des mutations compensatoires dans un des gènes codant pour une sous-unité de la gyrase. Ces mutations réduisent le niveau de surenroulement négatif du chromosome et permettent la croissance bactérienne. Une de ces mutations engendre la production d'une gyrase thermosensible. L’activité de surenroulement de la gyrase en absence de la topoisomérase I cause l’accumulation d’ADN hyper-surenroulé négativement à cause de la formation de R-loops. La surproduction de la RNase HI (rnhA), une enzyme qui dégrade l’ARN des R-loops, permet de prévenir l’accumulation d’un excès de surenroulement négatif. En absence de RNase HI, des R-loops sont aussi formés et peuvent être utilisés pour déclencher la réplication de l’ADN indépendamment du système normal oriC/DnaA, un phénomène connu sous le nom de « constitutive stable DNA replication » (cSDR). Pour mieux comprendre le lien entre la formation de R-loops et l’excès de surenroulement négatif, nous avons construit un mutant conditionnel topA rnhA gyrB(Ts) avec l’expression inductible de la RNase HI à partir d’un plasmide. Nous avons trouvé que l’ADN des cellules de ce mutant était excessivement relâché au lieu d'être hypersurenroulé négativement en conditions de pénurie de RNase HI. La relaxation de l’ADN a été montrée comme étant indépendante de l'activité de la topoisomérase IV. Les cellules du triple mutant topA rnhA gyrB(Ts) forment de très longs filaments remplis d’ADN, montrant ainsi un défaut de ségrégation des chromosomes. La surproduction de la topoisomérase III (topB), une enzyme qui peut effectuer la décaténation de l’ADN, a corrigé les problèmes de ségrégation sans toutefois restaurer le niveau de surenroulement de l’ADN. Nous avons constaté que des extraits protéiques du mutant topA rnhA gyrB(Ts) pouvaient inhiber l’activité de surenroulement négatif de la gyrase dans des extraits d’une souche sauvage, suggérant ainsi que la pénurie de RNase HI avait déclenché une réponse cellulaire d’inhibition de cette activité de la gyrase. De plus, des expériences in vivo et in vitro ont montré qu’en absence de RNase HI, l’activité ATP-dépendante de surenroulement négatif de la gyrase était inhibée, alors que l’activité ATP-indépendante de cette enzyme demeurait intacte. Des suppresseurs extragéniques du défaut de croissance du triple mutant topA rnhA gyrB(Ts) qui corrigent également les problèmes de surenroulement et de ségrégation des chromosomes ont pour la plupart été cartographiés dans des gènes impliqués dans la réplication de l’ADN, le métabolisme des R-loops, ou la formation de fimbriae. La deuxième partie de ce projet avait pour but de comprendre les rôles des topoisomérases de type IA (topoisomérase I et topoisomérase III) dans la ségrégation et la stabilité du génome de Escherichia coli. Pour étudier ces rôles, nous avons utilisé des approches de génétique combinées avec la cytométrie en flux, l’analyse de type Western blot et la microscopie. Nous avons constaté que le phénotype Par- et les défauts de ségrégation des chromosomes d’un mutant gyrB(Ts) avaient été corrigés en inactivant topA, mais uniquement en présence du gène topB. En outre, nous avons démontré que la surproduction de la topoisomérase III pouvait corriger le phénotype Par- du mutant gyrB(Ts) sans toutefois corriger les défauts de croissance de ce dernier. La surproduction de topoisomérase IV, enzyme responsable de la décaténation des chromosomes chez E. coli, ne pouvait pas remplacer la topoisomérase III. Nos résultats suggèrent que les topoisomérases de type IA jouent un rôle important dans la ségrégation des chromosomes lorsque la gyrase est inefficace. Pour étudier le rôle des topoisomérases de type IA dans la stabilité du génome, la troisième partie du projet, nous avons utilisé des approches génétiques combinées avec des tests de « spot » et la microscopie. Nous avons constaté que les cellules déficientes en topoisomérase I avaient des défauts de ségrégation de chromosomes et de croissance liés à un excès de surenroulement négatif, et que ces défauts pouvaient être corrigés en inactivant recQ, recA ou par la surproduction de la topoisomérase III. Le suppresseur extragénique oriC15::aph isolé dans la première partie du projet pouvait également corriger ces problèmes. Les cellules déficientes en topoisomérases de type IA formaient des très longs filaments remplis d’ADN d’apparence diffuse et réparti inégalement dans la cellule. Ces phénotypes pouvaient être partiellement corrigés par la surproduction de la RNase HI ou en inactivant recA, ou encore par des suppresseurs isolés dans la première partie du projet et impliques dans le cSDR (dnaT18::aph et rne59::aph). Donc, dans E. coli, les topoisomérases de type IA jouent un rôle dans la stabilité du génome en inhibant la réplication inappropriée à partir de oriC et de R-loops, et en empêchant les défauts de ségrégation liés à la recombinaison RecA-dépendante, par leur action avec RecQ. Les travaux rapportés ici révèlent que la réplication inappropriée et dérégulée est une source majeure de l’instabilité génomique. Empêcher la réplication inappropriée permet la ségrégation des chromosomes et le maintien d’un génome stable. La RNase HI et les topoisomérases de type IA jouent un rôle majeur dans la prévention de la réplication inappropriée. La RNase HI réalise cette tâche en modulant l’activité de surenroulement ATP-dependante de la gyrase, et en empêchant la réplication à partir des R-loops. Les topoisomérases de type IA assurent le maintien de la stabilité du génome en empêchant la réplication inappropriée à partir de oriC et des R-loops et en agissant avec RecQ pour résoudre des intermédiaires de recombinaison RecA-dépendants afin de permettre la ségrégation des chromosomes.

IA Critical Study of the Impact of Institutional Finance on Agriculture in Selected Villages of Ernakulam Revenue District

Vast changes have taken place in the field of institutional rural credit in India since the nationalisation of nineteen commercial banks in 1969. The supply of institutional finance to cultivators amounted to 63.2 percent of the total credit in 1981 compared to 31.2 percent in 1971. Insti tutionalisation of agricultural credit envisaged two objectives in general. One was to emancipate cultivators and farmers from the clutches of indigenous financiers and money lenders. The second was to make farmers financially capable of adopting the new technology or improved practices in agriculture so as to increase their agricultural production and thereby contributing to the development of agriculture in India. Though vast literature on Institutional Credi t and agriculture is available, no indepth and serious work examining thoroughly the cause of credit diversion has been undertaken so far. The present study is an attempt to fill up this gap. The study will be helpful to lending insti tutions, viz. Co-ope:r-atives, Commercial banks and various other insti tutional agencies in connection with their lending activity_ Also, the study will help government in .formulating proper policies that will insure a preferential treatment in favour of the most needy category of farmers and cultivators with respect to agricultural credit disbursement

THE PDETICAL WORKS DF GIRI-IA KUMAR MATHUR: SENSIBILITY AND TECHNIQUE

HINDI

FAIA : framework para la enseñanza de agentes en IA.

Resumen tomado del autor

Intervenci??n de una compa??ia de milicia alcarre??a en la batalla de Almansa de 1707

Se trata de un an??lisis sobre las distintas operaciones milicianas llevadas a cabo en la regi??n de Almansa a principios del siglo de las luces durante la guerra de sucesi??n espa??ola. Se analizan la importancia de la micro historia dentro del contexto global de la guerra y la importancia del voluntariado b??lico como componente esencial de la guerra de sucesi??n espa??ola y de otros conflictos armados anteriores a la Guerra Civil Espa??ola.

El último vuelo del condor : una experiencia colectiva de dramatización de textos.

En portada: De la crónica al teatro

O impacto da profissionalização na gestão de uma organização familiar : o caso da Condor S.A.

Esta pesquisa aborda o profissionalização de uma empresa familiar, situada no Planalto Norte de Santa Catarina, a Condor S.A., que em 1997, transferiu todos os executivos familiares para o Conselho de Administração, criado na ocasião para fiscalizar e dar suporte em termos de estratégia à nova diretoria executiva, não familiar, para gerir a empresa. O objetivo da pesquisa foi de identificar e avaliar as mudanças havidas nas estruturas e nos processos da Condor S.A. ocorridas após a profissionalização. A Condor é uma empresa familiar há mais de 70 anos. As empresas familiares têm seus pontos positivos, mas os negativos destacam-se quando a empresa começa a sentir os problemas de sobrevivência. As características próprias de cada empresa familiar, se não bem conduzidas afetarão a sobrevivência da empresa na atual e dinâmica competitividade. Assim, a importância de estudar as transformações possíveis numa empresa familiar antiga, da 3ª e 4ª gerações, frente à profissionalização da diretoria executiva e buscar as nuances ocorridas nesse processo. Pela análise da Condor S.A. concluiu-se que ocorreram diversas mudanças que contribuíram de forma positiva na gestão dessa empresa e que a forma escolhida para a profissionalização também se constituiu numa opção que está viabilizando a manutenção da empresa. Verificou-se, também, que os profissionais contratados que não conseguem atender aos anseios da empresa a curto prazo permanecem por pouco tempo. Esses profissionais, por pretenderem fazer uma mudança radical, não se atendo à cultura da empresa, normalmente não se entrosam na organização, e geram um fator de insucesso para a empresa e para a sua carreira. Dentre as principais mudanças organizacionais que ocorreram e que causaram impacto na sua gestão, pode-se destacar: a descentralização e a agilidade nas decisões; as metas e objetivos claros disseminados por toda a organização; maior transparência no contado da direção com todos os funcionários; a cobrança dos resultados pelas metas traçadas e não de forma subjetiva como ocorria anteriormente; a administração mais racional e objetiva e o planejamento estratégico.

Os desafios da internacionalização da indústria do varejo brasileiro de supermercados: o crescimento do Condor Super Center analisado em um estudo de caso para ensino

Este trabalho tem como objetivo oferecer um caso para ensino tendo como referência uma empresa brasileira do comércio varejista alimentício de supermercados e hipermercados. A estratégia de crescimento é o dilema central. Internacionalizar-se ou não? Atualmente, nenhuma empresa brasileira deste segmento se encontra internacionalizada por meio de lojas próprias no exterior. Uma visão histórica da empresa e de seu fundador é apresentada. Aspectos do setor, no Brasil, também são fornecidos. O caso coloca em evidência as características da organização e do seu presidente. Este trabalho apresenta mais do que um estudo de caso para ensino. Abordagens teóricas baseadas em conceitos de empreendedorismo, orientação empreendedora, empreendedorismo internacional e visão baseada em recursos são apresentadas como uma alternativa de análise. Uma contextualização introdutória é feita sobre a importância da internacionalização do varejo em relação à sociedade em rede. Uma comparação entre estudo de caso para pesquisa e um caso de ensino é oferecido no capítulo sobre metodologia. Disciplinas como gestão estratégica, empreendedorismo, internacionalização de empresas, teorias organizacionais, marketing e economia são boas opções para se empregar o caso proposto.

Distinctively variable sequence-based nuclear DNA markers for multilocus phylogeography of the soybean- and rice-infecting fungal pathogen Rhizoctonia solani AG-1 IA

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

"Eu pensei que ia sair daqui falando inglês": um estudo sistêmico-funcional sobre papéis sociais atribuídos por alunos de um curso de Letras/Inglês

Research in the area of teacher training in English as a Foreign Language (CELANI, 2003, 2004, 2010; PAIVA, 2000, 2003, 2005; VIEIRA-ABRAHÃO, 2010) articulates the complexity of beginning teachers classroom contexts aligned with teaching language as a social and professional practice of the teacher in training. To better understand this relationship, the present study is based on a corpus of transcribed interviews and questionnaires applied to 28 undergraduate students majoring in Letters/English emphasis, at a public university located in the interior of the Western Amazon region, soliciting their opinions about the reforms made in the curriculum of this Major. Interviews and questionnaires were used as data collection instruments to trace a profile of the students organized in Group 1, with freshmen and sophomore undergraduates who are following the 2009 curriculum, and Group 2, with junior and senior undergraduates who are following the 2006 curriculum. The objectives are to identify, to characterize and to analyze the types of pronouns, roles and social actors represented in the opinions of these students in relation to their teacher training curriculum. The theoretical support focuses on the challenge of historical and contemporary routes from English teachers initial education programs (MAGALHÃES; LIBERALLI, 2009; PAVAN; SILVA, 2010; ALVAREZ, 2010; VIANA, 2011; PAVAN, 2012). Our theoretical perspective is based on the Systemic Functional Grammar of Halliday (1994), Halliday and Hasan (1989), Halliday and Matthiessen (2004), Eggins (1994; 2004) and Thompson (2004). We focus on the concept of the Interpersonal meaning, specifically regarding the roles articulated in the studies by Delu (1991), Thompson and Thetela (1995), and in the Portuguese language such as Ramos (1997), Silva (2006) and Cabral (2009). Moreover, we ascribe van Leeuwen s (1997; 2003) theory of Representation of Social Actors as a theoretical framework in order to identify the sociological aspect of social actors represented in the students discourse. Within this scenario, the analysis unfolds on three levels: grammatical (pronouns), semantic (roles), and discursive (social actors). For the analysis of interpersonal realizations present in the students opinions, we use the computational program WordSmith Tools (SCOTT, 2010) and its applications Wordlist and Concord to quantify the occurrences of the pronouns I, You and They, which characterize the roles and social actors of the corpus. The results show that the students assigned the following roles to themselves: (i) apprentice to express their initial process of English language learning; (ii) freshman to reveal their choice of Major in Letters/English emphasis; (iii) future teacher to relate their expectations towards a practicing professional. To assign the roles to professors in the major, the students used the metaphor of modality (I think) to indicate the relationship of teacher training, while they are in the role of a student and as a future teacher. From these evidences the representation of the students as social actors emerges in roles such as: (i) active roles; (ii) passive roles and (iii) personalized roles. The social actors represented in the opinions of the students reflect the inclusion of these roles assigned to the actions expressed about their experiences and expectations derived from their teacher training classroom

Relação da imunoexpressão da bmp-2, bmpr-ia e bmpr-ii com o perfil clínico-patológico em carcinoma epidermóide de lábio inferior

Currently, bone morphogenetic proteins (BMPs) have effective participation in the growth of malignancies. Knowing that there are few studies involving BMPs and oral squamous cell carcinoma, this work constitutes an immunohistochemical study of BMP-2, BMPR IA and BMPR II in squamous cell carcinomas (SCC) of the lower lip relating to the clinical and pathological aspects of this lesion. The sample consisted of 40 cases of SCC of the lower lip, being 20 cases of SCC of the lower lip with regional metastasis and 20 cases without metastasis. We evaluated the intensity of expression (score 1 to mark absent / weak, score 2 for high ) and was found the percentage of labeled cells, where the score was 1 cases with 0 to 50% of positive cells, score 2 with 51 to 75% of positive cells, and score 3 more than 75% of positive cells. The sample comprised 72.5% of men with a mean age of 65.8 years, there was a predominance of stage II and 52.5% of the carcinomas were classified as low grade, being carcinoma with metastasis presenting most cases (70%) as carcinomas of high malignancy grade (p = 0.004). The largest number of cases of SCC of the lower lip that were in stages I / II (61, 9%) were classified as carcinomas of low grade malignancy and carcinomas in stages III / IV were classified as high-grade tumors (p = 0, 024). The BMP-2 showed strong intensity of immunostaining in 82.5%, BMPR-IA showed 55% of cases with an intensity of immunostaining absent / weak and BMPR-II showed 85% of cases with an intensity of immunostaining absent / weak. Only the protein BMPR-IA were significantly associated with all clinic-pathological parameters studied, metastasis (p <0.001), TNM (p <0.001) and histological grade of malignancy with (p = 0.028). The percentage of positive cells, all markers showed the highest number of cases with more than 75% of positive cells (score 3) and only BMPR-II showed statistical difference when related to the presence and absence of metastasis (p = 0.049 ). We conclude that there is disturbance in the BMP signaling pathway in EC-mediated lower lip and that high expression of BMP-2 associated with the expression of BMPR-IA and BMPR-II are associated with metastasis in carcinoma

Avaliação da expressão da BMP -2/4 e BMPR-IA em Carcinoma Epidermóide Oral metastático e não metastático

The expression of bone morphogenetic proteins (BMPs) is altered in a variety of human canceres. The BMP-2/4 and BMPR-IA were recently shown to be overexpressed in high-risk premalignant and malignant lesions of oral epithelium. The present study analysed the expression of BMP-2/4 and BMPR-IA in Oral Squamous Cell Carcinoma (OSCC) such as their implications in disease prognostic using munohistochemistry. Ten cases of Oral Fibroepithelial Hiperplasia were selected as a control group. The experimental group included 16 cases of OSCC without metastases and 7 cases of OSCC metastatic. The presence or absence of nodal metastases was used as parameter to evaluated the disease prognostic. The results demonstrated weak immunoreactivity for BMP-2/4 and BMPR-IA in every case of the control group. In the cases of OSCC with metastases an overexpression of BMP-2/4 (71,4%) was observed while the BMPR-IA showed weak expression (85,7%). In the cases of OSCC without metastases BMP-2/4 (62,5%) and BMPR-IA showed strong immunostaining standing out an overexpression of the receptor in all the specimens. Observed statistical significance for correlation between the oral cancer prognostic and the staining intensity of the BMP-2/4 (p=0,002). There wasn t statistical significance for association between the staining intensity of the BMPR-IA and the disease prognostic (p<0,001). In conclusion, this findings suggest that the overexpression of BMP-2/4 associated with the loss of expression of the BMPR-IA in OSCC metastatic has prognostic relevance, as the loss of sensitivity to BMPs can be an indicative of metastases development in OSCC

Estudo clínico-patológico do carcinoma epidermóide de língua e imunoistoquímico das proteínas BMP-2, BMPR-IA, BMPR-II e endoglina

Bone morphogenetic proteins (BMPs) are cytokines involved in proliferation and angiogenesis of many kind of human cancer. The present study analyzed the immunohistochemical expression of BMP-2, BMPR-II, BMPR-IA and endoglin (CD105) and their relationship with the biological behavior and local angiogenesis in tongue oral squamous cells carcinoma (SCC). The sample consisted of 25 cases of tongue SCC without metastasis, 25 tongue SCC with metastasis and 25 cases of Inflamatory Fibrous Hyperplasia (IFH).The histological grade of malignancy proposed by Bryne (1998), adapted by Miranda (2002) was used to classify all tongue SCC cases. Score 0 was attributed to absent-weak immunoexpression and score 1 for strong immunostaning and pattern of distribution was focal or diffuse. Microvessel counts (MVC) was established for CD105. Most of the patients with tongue SCC was male. The principal age in tongue SCC without metastasis was over 65 years and in tongue SCC with metastasis was between 45-65 years. There were predominance of stage II in TNM and in the specimens with high-grade, independent of studied group. For BMP-2, 56% of tongue SCC without metastasis and 72% tongue SCC with metastasis exhibited score 1 while the IFH showed secore 0 in 72% of the cases, with statistical association (p=0,007). Considering the BMPR-II, 52% of tongue SCC without metastasis exhibited score 0; 56% tongue SCC with metastasis and 60% IFH showed score 1. The majority cases of BMPR-IA demonstrated score 1 and 100% of CD105 exhibited strong immunoexpression in tongue SCC. Regarding the pattern distribution, it was noted a tendency to diffuse pattern for the proteins in all groups. The means of MVC were similar in tongue SCC without metastasis (32,91) and in tongue SCC with metastasis (32,05), however existed statistical difference with IFH (p<0,001). There was statistical association of BMP-2 expression with BMPR-II (p=0,008), BMPR-IA (p=0,006) and CD105 (p=0,046). An association between TNM and BMP-2 immunoexpression and their receptors was not detected, nevertheless this association was found with MVC (p=0,047) whose averages were higher for the stages II (35,97) e IV (35,69). No association between histological grading and these proteins was observed. This study suggests that the superexpression of BMP-2 signaling pathways acts on cell proliferation in tongue SCC and can be implicated with more invasive potential. Additionaly, the CD105 is a potent biological marker of neovascularization in this neoplasm and their association with BMP-2 and BMPR-IA receptor, showed that this type of cancer in BMP-2 is presented as pro-angiogenic in the metastatic process