The effects of the environment on the stability and expression of genetically engineered plasmids

The lac promoter is widely used in plasmid expression systems, even though it is prone to catabolite repression. As a consequence glycerol is often used as an alternative carbon source. Three plasmids containing various sizes of the staphylococcal protein A (SPA) gene, which are under the control of the lac promoter were investigated in continuous culture, to evaluate the effects of nutrient limitations on their stability and expression. The fears of catabolite repression were dispelled as a low expression plasmid (pPA16) produced a greater amount of truncated SPA under glucose limiting conditions (11 ug mg-1 cell protein) when compared to that using glycerol (8 ug mg-1 cell protein). Segregational instability was also observed under glycerol limiting conditions at all the dilution rates investigated. Whereas pPA16 was relatively stable under glucose limiting conditions, with SPA production being continuous. Experiments using excess glycerol with limited ammonium increased the stability of pPA16, (when compared to limited glycerol) with expression of SPA being continuous but reduced (6 ug mg-1 cell protein). With excess glucose and limited ammonium the copy numbers remained high but expression of SPA paralled that produced under glucose limiting conditions. This might indicate that the higher levels of glucose are reducing expression (catabolite repression) or that the low level of ammonium is affecting protein production. A high expression plasmid (pPA31) produced nearly 100 ug full length SPA mg-1 cell protein, while another high expression plasmid (pPA34) producing truncated SPA proved to be very unstable. An ELISA was developed to detect the SPA produced by these experiments, which could be adapted for western blotting or immunogold probing using electron microscopy. SPA was localised in electron lucent areas present in the periplasmic space of the E. coli host harbouring pPA16. While in the same host containing pPA31, SPA was localised not only in electron lucent areas but also around the whole of the outer-membrane.

Finance and entrepreneurial entry

This paper investigates the determinants of both the volume of finance and the sources of finance of business start-ups in 42 countries. Using the Global Entrepreneurship Monitor (GEM) surveys for 1998-2003, we jointly examine how the institutional business environment and individual characteristics of entrepreneurs affect the financing of entrepreneurs. We find that the property rights system is the most significant determinant of both the total volume of finance and of the use of external finance for the individual start-up project. In addition, our findings suggest that that the supply of informal finance is associated with the higher share of external sources in start-up finance, whereas the size of the formal financial sector appears to play a more important role for the volume of entrepreneurial self-finance. Our results suggest that the use of external finance by start-ups correlates with the extent of financial restrictions in a country in a non-monotonic way. We find that some financial restrictions may enhance the use of external funds by entrepreneurs, yet the sign of this effect reverses and the coefficient becomes stronger for the high level of financial repression by the government. Finally, our findings imply that the characteristics of the start-up finance are also affected by various individual characteristics of entrepreneurs, their growth intentions and ownership structure.

Diesseits und jenseits der Oder:Judith Hermanns literarische Auseinandersetzung mit Theodor Fontane in Sommerhaus, später

Judith Hermann's works have attracted considerable criticism for their supposedly slight portrayal of passively drifting characters and for their alleged failure to engage with the socio-political realities of contemporary life in the Berlin Republic. Only very recently have scholars paid attention to the hidden concern with memory expressed in her books, and have set out to examine their intertextual depth. This paper explores these previously neglected historical references in Summerhouse, later and analyses the book's intricate intertextual allusions with specific reference to Theodor Fontane's works. It examines how the tentative existence, which Hermann's characters experience, is the product of a hesitant and fruitless confrontation with questions of German history and nationhood. Using pervasive water imagery, Hermann shows present-day Germany as a continually contested territory with a fluid identity shaped by an abundance of conflicting narratives. In this context, the allusions to Fontane as a representative of the Wilhelminian period serve as references to a continuing German tradition of repression and marginalisation. At the same time, Hermann recognises Fontane's ambivalent political stance combining elements of social criticism with a general endorsement of social order. Ultimately, the seemingly indifferent attitude of Hermann's characters and the elegiac style used to portray them, emerge as a distancing mechanism that functions as a postmodern variant of Fontane's irony and is shaped by a similar sense of skepticism towards developments in German society and national history. © 2012 Springer Science+Business Media B.V.

Pichia pastoris regulates its gene-specific response to different carbon sources at the transcriptional, rather than the translational, level

Background: The methylotrophic, Crabtree-negative yeast Pichia pastoris is widely used as a heterologous protein production host. Strong inducible promoters derived from methanol utilization genes or constitutive glycolytic promoters are typically used to drive gene expression. Notably, genes involved in methanol utilization are not only repressed by the presence of glucose, but also by glycerol. This unusual regulatory behavior prompted us to study the regulation of carbon substrate utilization in different bioprocess conditions on a genome wide scale. Results: We performed microarray analysis on the total mRNA population as well as mRNA that had been fractionated according to ribosome occupancy. Translationally quiescent mRNAs were defined as being associated with single ribosomes (monosomes) and highly-translated mRNAs with multiple ribosomes (polysomes). We found that despite their lower growth rates, global translation was most active in methanol-grown P. pastoris cells, followed by excess glycerol- or glucose-grown cells. Transcript-specific translational responses were found to be minimal, while extensive transcriptional regulation was observed for cells grown on different carbon sources. Due to their respiratory metabolism, cells grown in excess glucose or glycerol had very similar expression profiles. Genes subject to glucose repression were mainly involved in the metabolism of alternative carbon sources including the control of glycerol uptake and metabolism. Peroxisomal and methanol utilization genes were confirmed to be subject to carbon substrate repression in excess glucose or glycerol, but were found to be strongly de-repressed in limiting glucose-conditions (as are often applied in fed batch cultivations) in addition to induction by methanol. Conclusions: P. pastoris cells grown in excess glycerol or glucose have similar transcript profiles in contrast to S. cerevisiae cells, in which the transcriptional response to these carbon sources is very different. The main response to different growth conditions in P. pastoris is transcriptional; translational regulation was not transcript-specific. The high proportion of mRNAs associated with polysomes in methanol-grown cells is a major finding of this study; it reveals that high productivity during methanol induction is directly linked to the growth condition and not only to promoter strength.

REST is a hypoxia-responsive transcriptional repressor

Cellular exposure to hypoxia results in altered gene expression in a range of physiologic and pathophysiologic states. Discrete cohorts of genes can be either up- or down-regulated in response to hypoxia. While the Hypoxia-Inducible Factor (HIF) is the primary driver of hypoxia-induced adaptive gene expression, less is known about the signalling mechanisms regulating hypoxiadependent gene repression. Using RNA-seq, we demonstrate that equivalent numbers of genes are induced and repressed in human embryonic kidney (HEK293) cells. We demonstrate that nuclear localization of the Repressor Element 1-Silencing Transcription factor (REST) is induced in hypoxia and that REST is responsible for regulating approximately 20% of the hypoxia-repressed genes. Using chromatin immunoprecipitation assays we demonstrate that REST-dependent gene repression is at least in part mediated by direct binding to the promoters of target genes. Based on these data, we propose that REST is a key mediator of gene repression in hypoxia.

Literatura femenina en el Peru decimononico. La cuestion del naturalismo y el feminismo en la obra de Mercedes Cabello de Carbonera

Women writers in the nineteenth century were often underestimated and in some cases completely ignored. At the end of that century, a considerable group of Peruvian women writers had a significant influence in the development of Lima's cultural life. Either together in the “veladas literarias” or individually in their own work, they showed a common interest in women's concerns and especially in the problems regarding women education or, better to say, the lack of it. Although frequently these writers just followed the paths men have marked for them, they often tried to find their own ways of expression in their works. ^ This dissertation examined the cultural life in Lima at the end of the century and concentrated in one of these writers, Mercedes Cabello de Carbonera (1845–1909), whose work was analyzed from a feminist point of view. American critics had been chosen for the analysis and especially the feminist theories of Sandra Gilbert, Susan Gubar and Elaine Showalter. They were applied to three of Cabello de Carbonera's novels: Sacrificio y recompensa, Las consecuencias and Blanca Sol. ^ The study attempted to prove how often women opinions were distorted by the male tradition and how they tried a different way of expression through the metaphors and symbols referring to the state of repression women were in. Beneath the surface of their work lies a determined feminine consciousness. ^

La represión sexual como tema recurrente en la obra de Federico García Lorca

The purpose of this dissertation is to demonstrate that sexual repression is a recurring theme throughout a selection of works by Federico García Lorca. The introductory chapter focuses on the sexual theories of Sigmund Freud and Michel Foucault in setting the foundation for an analysis of sexually frustrated characters. In chapter two, an analysis of Lorca's rural trilogy reveals how marginalized female characters struggle to preserve societal customs that subjugate them to the patriarchal hegemony by limiting their free will and their sexual freedom. The subsequent chapter elaborates on the theme of repression in poems where the oppressed voice expresses and denounces the intolerance and persecution of those who condemn homosexuality through metaphors of darkness, impurity, and barren love. Chapter four analyzes two avant-garde plays where the underlying message is that of a society that hampers the expression of homosexual love by silencing desire and forcing individuals to mask their identities. The conclusion demonstrates once more the effects of society on sexual freedom, as well as explaining that the recurrence of repressed women is not a pretext in portraying the homosexual saga, but a pre-text in presenting them as an equally marginalized group.

Representation of people of color in the nineteenth century slavery novel in Cuba and Brazil

This dissertation explores the similarities and differences which characterize the depiction of people of color in certain representative nineteenth century Cuban and Brazilian slavery novels as a function of the authorial approach of each territory's literary tradition toward the issues of slavery, racial prejudice, and people of color. The selected texts, derived from the peak periods in slavery literature of each territory, include Francisco , by Anselmo Snárez y Romero; Sab, by Gertrudis Gómez de Avellaneda; Cecilia Valdés , by Cirilo Villaverde; A escrava Isaura, by Bernardo Guimarães; O mulato, by Aluísio Azevedo; and Bom-Crioulo, by Adolfo Caminha. While the present study explores the enslavement, abuse, and discrimination of people of color as a consequence of a deep-seated discourse of power, privilege and racial superiority, it focuses more extensively on the representation of people of color, particularly in their capacity to constructively appropriate the cultural values of the white dominant group and recognize their identity as ambiguous. ^ Said's theories of Orientalist discourse and geography and formation as well as Dube's perspective on subaltern-oriented studies provide a theoretical framework for exploring the response of slavery writers whose common exposure to slavery but dissimilar socio-political contexts generate some startling findings. Crafted within a period of political repression, fear of black revolt, factional in-fighting as well as strong socioeconomic ties to the slaveholding class, the Cuban texts generally fashioned an approach to slavery as one marked by moderation, reform, and cultural counter discourse and consequently depict people of color with a more passive but culturally authentic outlook. On the other hand, the Brazilian response to the issue of slavery, steeped in an ideological amalgam of liberalism, positivism, republicanism, and abolitionism, is characterized by overt opposition to slavery and a representation of people of color that is less concerned with cross-cultural input but reclaims their humanity as highly educable and socially mobile persons in search of greater freedoms. Ultimately, there is a shared message of higher significance couched in the worthwhile mission of raising slaves to the level of men. ^

Bolivia's Uncertain Future

Edited by former LACC Director, Eduardo Gamarra, this issue asked prominent Bolivian journalists and social scientists to critically analyze the first year and a half of Evo Morales’ government. Popularly elected in December 2005, Morales promised to conduct a revolution in democracy. In this collection of essays, the objective is to show a different view than the image of Morales as the Bolivian Nelson Mandela who freed his indigenous brethren from repression. The essays gathered here tell the story about how Bolivia’s first indigenous president has attempted to change Bolivia. These essays show that Morales’ first 18 months in office have been filled with promise, controversy, and conflict.

Global Capitalism, Immigrant Labor, and the Struggle for Justice

Around the world borders are militarized, states are stepping up repressive anti-immigrant controls, and native publics are turning immigrants into scapegoats for the spiraling crisis of global capitalism. The massive displacement and primitive accumulation unleashed by free trade agreements and neo-liberal policies, as well as state and “private” violence has resulted in a virtually inexhaustible immigrant labor reserve for the global economy. State controls over immigration and immigrant labor have several functions for the system: 1) state repression and criminalization of undocumented immigration make immigrants vulnerable and deportable and therefore subject to conditions of super-exploitation, super-control and hyper-surveillance; 2) anti-immigrant repressive apparatuses are themselves ever more important sources of accumulation, ranging from private for-profit immigrant detention centers, to the militarization of borders, and the purchase by states of military hardware and systems of surveillance. Immigrant labor is extremely profitable for the transnational corporate economy; 3) the anti-immigrant policies associated with repressive state apparatuses help turn attention away from the crisis of global capitalism among more privileged sectors of the working class and convert immigrant workers into scapegoats for the crisis, thus deflecting attention from the root causes of the crisis and undermining working class unity. This article focuses on structural and historical underpinnings of the phenomenon of immigrant labor in the new global capitalist system and on how the rise of a globally integrated production and financial system, a transnational capitalist class, and transnational state apparatuses, have led to a reorganization of the world market in labor, including deeper reliance on a rapidly expanding reserve army of immigrant labor and a vicious new anti-immigrant politics. It looks at the United States as an illustration of the larger worldwide situation with regard to immigration and immigrant justice. Finally, it explores the rise of an immigrant justice movement around the world, observes the leading role that immigrant workers often play in worker’s struggles and that a mass immigrant rights movement is at the cutting edge of the struggle against transnational corporate exploitation. We call for replacing the whole concept of national citizenship with that of global citizenship as the only rallying cry that can assure justice and equality for all.

A Body Outside the Kremlin

A BODY OUTSIDE THE KREMLIN is a historical mystery novel set in the Northern Camps of Special Significance, a Soviet Russian penal institution based in the Solovetsky Archipelago during the 1920s. The protagonist, working first with the camp authorities, then in spite of their disapproval, solves the murder of a fellow prisoner. In the process he improves his position within the camp, while also becoming hardened to the brutal necessities of camp life. Prior to the establishment of the penal camp, the Solovetsky Archipelago was the site of an important Russian Orthodox monastery, and the mystery proves to involve valuables, particularly icons, seized from the monks by the Soviet secret police. Thus the novel treats themes not only of statist repression, but also religious epiphany and the problems of true perception in a world of symbols.

Punição e controle: o “caminho punitivo” do jovem autor de ato infracional em Fortaleza - CE

This research aims to analyze the punitive pathway taken by youth for committing acts of offense in Fortaleza, capital city of Ceará, in Brazil. Therefore, we focus the analysis in institutions that mark the beginning of punitive “institutionalization” of youth in the city, such as the Child and Adolescent Specialized Police Precinct, the Luis Barros Montenegro Shelter Unit, the Public Prosecutor’s Office for Childhood and Youth, and the Child and Youth Court. Ethnography and semi-structured interviews were used as methodological tools to approach the research subjects and relevant places for the research, seeking to highlight their punitive perspective. As a result, we find that the punishment and control imposed in such loci are an extension of the punishment and control used against these same youths in society. Considering that the analyzed institutions exist in society and are composed by its members, they do not surpass the perspective of repression, control and punishment carried out towards a segment of the population, especially towards those that Souza (2011; 2012) called “subcitizens”.

A inserção da família no processo socioeducativo de adolescentes em privação de liberdade

This master thesis has the main goal investigate how families are inserted in the socioeducational process of teenagers who are undergoing social measures of liberty deprivation. The specific objectives are: to characterize the family´s living together of adolescents deprived of their freedom and their families, from the actions and routines of the socio-educational system; to assess the professional working links in the context of socio-education, in order to care and strengthening families of the adolescents; to investigate how families evaluate the operation of socio-educational process in which adolescents are met. Method: to achieve the proposed objectives, data collection occurred in complementary steps: the first phase took place from visits to socioeducational units of liberty deprivation of RN, and dialogues with professionals working in socio-education. Subsequently, action research stage was carried out, from the insertion of the researcher in the extension project Family and the struggle for the effectiveness of the National Socio Service System, that aimed to strengthen monitoring the adolescents socio-education by their families, and had as methodology the conversation circles and thematic workshops. Lastly, were performed reading and analysis of the references to the family in the Individual Care Plans (PIAs) for adolescents. The information gathered was recorded in field diaries and subjected to thematic content analysis. This research was guided by the Marxist theoretical framework, structured on the understanding of the involvement of adolescents with illegal acts as a development and expression of the social question. From this theoretical framework, the prevailing view in the capitalist society of adolescents in conflict with the law as individuals who are treated by means of repression and segregation and the weakness of social policies is questioned, both in the execution of their own socioeducational measures as the articulation of network services for adolescent protection and strengthening your family. Results: in relation to the operation of the socioeducational system in RN in general, it was observed a state of unhealthy physical spaces and institutional practices that violate human rights, idleness and lack of access to social rights, and criminalization and institutionalization of poor young people and their families. With regard to family´s living together, it was noticed great distance between principles and guidelines recommended by the SINASE, about acquaintanceship and family strengthening, and every day practices of socio-education in RN: serious violations were observed that undermine the family´s living together, as the distance between the socio-educational units of deprivation of liberty and the cities where families live; absence, irregularity and poor conditions in carrying out the family visits; lack of conjugal visits; restricted and unarticulated actions for the care and strengthening of the families of adolescents, most of whom live in poverty or extreme poverty. Finally, it was found a number of blamefully and punishments to the family, including practices such as inward inspection (visual inspection while naked and squant), plus a series of violence and omissions care that sick family members and weaken the links between adolescents and their families.

Relaciones de poder y guerra

Este artículo introduce los conceptos de relaciones de poder analizados por Michel Foucault, haciendo hincapié en el vínculo del ejercicio del poder en las relaciones internacionales. Por ello, se analizan el marco de relaciones que el ejercicio de dicho poder genera, así como las tipologías de relaciones entre Estados, que igualmente genera el poder. El artículo presenta los paradigmas de las relaciones internacionales y su evolución en el tiempo, estableciendo puentes con las consideraciones de Foucault con respecto al poder, la guerra, la represión o las razas. En este sentido, el concepto de guerra de razas se antoja fundamental para identificar dicha noción como la versión más extrema del paradigma realista. Dicha guerra de razas no constituye una categoría que integre necesariamente el concepto biológico, sino que se refiere al proceso de imposición de la identidad del grupo dominante, la cual puede contener aspectos étnicos, socioculturales y políticos. No obstante, ni el realismo ni su visión extrema son los únicos paradigmas existentes, ya que la evolución de los paradigmas alternativos ha conllevado la génesis del reflectivismo como alternativa al realismo.

El PCE y el movimiento comunista internacional (1969-1977)

El artículo se centra en la actividad internacional del Partido Comunista de España (PCE) desde las secuelas de la represión de la Primavera de Praga hasta 1977. Fue un período caracterizado por el principio del declive definitivo del movimiento comunista internacional. El análisis abarca múltiples cuestiones. Examina las crecientes críticas del PCE hacia el modelo soviético, y sus repercusiones en las relaciones entre el partido liderado por Santiago Carrillo y el movimiento comunista. Además, el artículo explora el intento del PCE de promover un nuevo tipo de internacionalismo en Europa occidental, auspiciando una renovada colaboración entre comunistas y socialistas y aprovechando las condiciones brindadas por la distensión. En este contexto tuvo lugar el surgimiento del eurocomunismo, que el ensayo analiza ilustrando los factores internos y externos que determinaron su crisis en la segunda mitad de los setenta.