The Social Institution and Inscription of Child Marriage in the Terai Region of Nepal

Marriage is typically considered to be one of the foundations of the family unit, and family is thought to be one of the pillars of society. Because marriage is such a primary social concept, its forms and functions may be taken for granted, despite the fact that it is culturally dependent. For example, the idea that one must be in love in order to get married may be more cultural ideal than universal idea—and is thus not true of many cultures. Arranged marriages are still performed in many parts of the world, and can be means of building kinship circles and economic ties, rather than serving as declarations of cultivated, romantic love. While the reasons for marriage may be negotiable, certain human rights regarding marriage are not. In some regions, including South Asia, marriages are often arranged between children, a status typically defined as people under the age of eighteen. In Nepal, the numbers of girls married off before they become legal adults, or before they even hit puberty are still fairly staggering, despite the practice being declared illegal since 1963, more than 50 years ago. Certain arguments in favor of child brides are that it is an important cultural practice, or that a girl is safer from sexual predators if she is married. However, the reality of premature marriages is that the girls are subject to many health issues, including psychological ones, lose prospects of education, and often times, the husbands prove to be their sexual assaulters anyway. Further problematizing the practice is the fact that not all children are equally at risk. Girls do typically marry younger than boys, though both genders are susceptible to child marriage. If a girl is from a particular region, religion, ethnic group, or caste, her probability of being married early will also rise, often because of economic considerations of the dowry. Considering the myriad factors that influence the practice of child marriage in Nepal, as well as the proven inadequacy of the law, it is important to locate and frame the practice within the cultures in which it exists. With that said, one must question where the practice exists the most, why it exists there the most, and to what extent the practice of child marriage is a symptom of a larger dynamic of gender inequality. After spending time in the Terai, where the practice is the most common, it is my opinion that child marriage would not persist if it were not for the pervasiveness of practices such as the dowry, the unequal emphasis of marriage for women, and the practice of the daughter moving into the husband’s family’s home, all of which strengthen the existing patriarchy and prevent local girls from becoming self-determined

Functional comparison of S/MARs ('scaffold/matrix attachment regions') and insulators in a chromosomal context

Im eukaryontischen Zellkern ist die DNA durch die Assoziation mit Histonen als Chromatin organisiert. Ein gängiges Modell der Organisation des Interphasechromatins ist die Ausbildung einer Schlaufenstruktur in unabhängig regulierte Gendomänen. Diese werden durch S/MAR-Elemente (‘scaffold/matrix attachment regions’) begrenzt, welche durch ihre Bindung an die Kernmatrix die Basis der Chromatinschlaufen bilden. S/MAR-Elemente besitzen jedoch nicht nur strukturgebende Funktion, sondern sind auch entscheidend an der Genregulation beteiligt. In der vorliegenden Arbeit wurde der Einfluss dieser Elemente auf die Expressionseigenschaften von Transgenen in den Zelllinien CHO K1 und NIH3T3 mit verschiedenen Untersuchungssystemen analysiert. Eine Analyse in Klongemischen zeigte eine deutliche transkriptionsaktivierende Wirkung dieser Elemente. Da der genomische Integrationsort einen nicht vorhersagbaren Einfluss auf die Transgenexpression hat, wurde im folgenden untersucht, ob die S/MAR-Wirkung dominant ist oder ebenfalls vom Integrationsort abhängt. Hierzu wurde ein auf sequenzspezifischer Exzision beruhender Ansatz genutzt. Dieser zeigte eine vom Integrationsort abhängige aktivierende oder reprimierende sowie von Chromatineinflüssen abschirmende Wirkung dieser Elemente. Daher wurde im dritten Teil der Arbeit die S/MAR-Wirkung mit der von Chromatin-Insulatoren verglichen, welche ebenfalls eine positionsunabhängige Genexpression ermöglichen. Zu diesem Zweck wurde ein auf Rekombinase- vermitteltem Kassettenaustausch basierendes Konzept entwickelt, welches den Vergleich der S/MAR- und Insulator-Wirkung sowohl in zufälligen Integrationsorten als auch vor dem gleichen genomischen Hintergrund erlaubt. In zufälligen Integrationsorten wurde ein aktivierender und von der genomischen Umgebung abschirmender Einfluss dieser Elemente bestätigt. Jedoch wurde deutlich, dass ihre Wirkung stark kontextabhängig ist.In the eukaryotic nucleus the DNA is associated with histones and like this organized as chromatin. A current model of the organization of interphase chromatin is the formation of a loop structure of independently regulated gene domains. These are bordered by S/MAR elements (‘scaffold/matrix attachment regions’) which build up the basis of chromatin loops by binding to the nuclear matrix. S/MAR elements not only possess structural function but are also crucially involved in gene regulation. In this work the influence of these elements on expression properties of transgenes was analysed in the cell lines CHO K1 and NIH 3T3 using different examination systems. An analysis of clone mixtures showed a significant transcription activating influence of these elements. As the genomic integration site exerts a non-predictable influence on transgene expression, in the following it was examined if the S/MAR effect is dominant or depends also on the site of integration. To this end an approach based on site-specific excision was used. It was shown that S/MAR elements exert an activating or repressing effect depending on the site of integration and protect against influences of neighbouring chromatin. Therefore in the third part of the work the S/MAR effect was compared to the effect of chromatin insulators which also confer position-independent gene expression. For this a concept making use of recombinase-mediated cassette exchange was developed, which allows the comparison of S/MAR and insulator effects in random integration loci and also in the same genomic surrounding. In random integration loci it was confirmed that these elements have an activating influence and protect against position-effects. But it became clear that the effects are strongly context-dependent. As cassette-exchange was inefficient due to a high spontaneous resistance rate of the cell line, strategies for improving efficiency were developed

Cell lines development through targeted integration involving a CRISPR-Mad7-based system in CHO-DG44

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Recombinant protein expression by targeting pre-selected chromosomal loci

<p>Abstract</p> <p>Background</p> <p>Recombinant protein expression in mammalian cells is mostly achieved by stable integration of transgenes into the chromosomal DNA of established cell lines. The chromosomal surroundings have strong influences on the expression of transgenes. The exploitation of defined loci by targeting expression constructs with different regulatory elements is an approach to design high level expression systems. Further, this allows to evaluate the impact of chromosomal surroundings on distinct vector constructs.</p> <p>Results</p> <p>We explored antibody expression upon targeting diverse expression constructs into previously tagged loci in CHO-K1 and HEK293 cells that exhibit high reporter gene expression. These loci were selected by random transfer of reporter cassettes and subsequent screening. Both, retroviral infection and plasmid transfection with eGFP or antibody expression cassettes were employed for tagging. The tagged cell clones were screened for expression and single copy integration. Cell clones producing > 20 pg/cell in 24 hours could be identified. Selected integration sites that had been flanked with heterologous recombinase target sites (FRTs) were targeted by Flp recombinase mediated cassette exchange (RMCE). The results give proof of principle for consistent protein expression upon RMCE. Upon targeting antibody expression cassettes 90-100% of all resulting cell clones showed correct integration. Antibody production was found to be highly consistent within the individual cell clones as expected from their isogenic nature. However, the nature and orientation of expression control elements revealed to be critical. The impact of different promoters was examined with the tag-and-targeting approach. For each of the chosen promoters high expression sites were identified. However, each site supported the chosen promoters to a different extent, indicating that the strength of a particular promoter is dominantly defined by its chromosomal context.</p> <p>Conclusion</p> <p>RMCE provides a powerful method to specifically design vectors for optimized gene expression with high accuracy. Upon considering the specific requirements of chromosomal sites this method provides a unique tool to exploit such sites for predictable expression of biotechnologically relevant proteins such as antibodies.</p

Educação ambiental no IPBeja - exemplos de eco-atividades desenvolvidas com os estudantes

O programa Eco-escolas tem vindo a ser reconhecido pelo potencial que as eco-escolas representam no caminho para uma educação ambiental orientada para a sustentabilidade. Temos verificado que a adesão tem vindo a aumentar, bem como o número de atividades incluídas no programa. Espera-se que, iniciando-se na sala de aula, as actividades se expandam a toda a escola e possam vir a promover a mudança na comunidade em geral. Na Escola Superior Agrária do Instituto Politécnico de Beja (ESA/IPBEJA) as atividades têm sido desenvolvidas nos diferentes eixos que o programa contempla, sendo a conservação da Biodiversidade e o esforço para a Diminuição da utilização dos plásticos dois temas desenvolvidos no âmbito de duas unidades curriculares das formações em Agronomia e em Tecnologia dos alimentos. Relativamente à Biodiversidade, vamos dar continuidade a alguns trabalhos que temos vindo a desenvolver em sala de aula, acreditando ser possível divulgar e envolver toda a comunidade no sentido de (i) avaliar o estado dos espaços verdes do Campus do IPBeja, com base em bioindicadores e (ii) monitorizar a fauna auxiliar das culturas do Centro Hortofrutícola. Pretende-se promover a conservação dessas espécies e divulgar todo o seu contributo para a limitação natural das pragas existentes nestas culturas. Com relação à Diminuição da utilização dos plásticos no âmbito da Tecnologia dos alimentos, procura-se o desenvolvimento de trabalhos pelos alunos que visam a criação de novas embalagens biodegradáveis (bioplásticos e comestíveis) que substituam as plásticas existentes no mercado. Para além do trabalho dos conteúdos das Unidades Curriculares, estas atividades permitem sensibilizar para as problemáticas ambientais que integram o programa Eco-escolas beneficiando a comunidade local e a sociedade em geral. Assim, elas são antes de mais, atividades de Educação Ambiental

The developmental origin of adolescent alcohol use: Findings from the Mater University Study of Pregnancy and its outcomes

Background It is unclear whether fetal alcohol exposure contributes to alcohol use in adolescence. In this study, we examine the association between maternal alcohol use in pregnancy and adolescents’ drinking patterns at age 14. Methods The association of maternal alcohol exposure with early drinking was examined in 4363 adolescents taking part to the Mater University Study of Pregnancy (MUSP) and its outcomes, a population based birth cohort study commenced in Brisbane (Australia) in 1981. Mothers and children were followed up at birth, 5 and 14 years after the initial interview. Maternal alcohol use was assessed before and during pregnancy and at the 5 years follow-up. Adolescents’ alcohol use was assessed at child age 14. Results In multivariable analysis those born of mothers who consumed 3+ glasses during pregnancy were at increased risk to report drinking 3+ glasses compared with those whose mothers reported no drinking or drinking up to 2 glasses. Comparisons controlling for drinking before pregnancy and at age 5 found the averaged odds ratio of maternal drinking in pregnancy on risk of reporting alcohol consumption of 3 and more glasses at age 14 was 2.74 (CI 1.70, 4.22). Conclusion Our study suggests that they maybe a biological origin of early drinking. Further studies are needed to better disentangle the nature of the association and the role of other possible confounding factors

Analysis of ligation and DNA binding by Escherichia coli DNA ligase (LigA).

NAD+-dependent DNA ligases are essential enzymes in bacteria, with the most widely studied of this class of enzymes being LigA from Escherichia coli. NAD+-dependent DNA ligases comprise several discrete structural domains, including a BRCT domain at the C-terminus that is highly-conserved in this group of proteins. The over-expression and purification of various fragments of E. coli LigA allowed the investigation of the different domains in DNA-binding and ligation by this enzyme. Compared to the full-length protein, the deletion of the BRCT domain from LigA reduced in vitro ligation activity by 3-fold and also reduced DNA binding. Using an E. coli strain harbouring a temperature-sensitive mutation of ligA, the over-expression of protein with its BRCT domain deleted enabled growth at the non-permissive temperature. In gel-mobility shift experiments, the isolated BRCT domain bound DNA in a stable manner and to a wider range of DNA molecules compared to full LigA. Thus, the BRCT domain of E. coli LigA can bind DNA, but it is not essential for DNA nick-joining activity in vitro or in vivo

Photodissociation dynamics of the HCNN radical

The photodissociation dynamics of the diazomethyl (HCNN) radical have been studied using fast radical beam photofragment translational spectroscopy. A photofragment yield spectrum was obtained for the range of 25 510-40 820 cm-1, and photodissociation was shown to occur for energies above 25 600 cm-1. The only product channel observed was the formation of CH and N2. Fragment translational energy and angular distributions were obtained at several energies in the range covered by the photofragment yield spectrum. The fragment translational energy distributions showed at least two distinct features at energies up to 4.59 eV, and were not well fit by phase space theory at any of the excitation energies studied. A revised C-N bond dissociation energy and heat of formation for HCNN, D0 (HC-NN) =1.139±0.019 eV and Δf H0 (HCNN) =5.010±0.023 eV, were determined. © 2006 American Institute of Physics.Ann Elise Faulhaber, Jason R. Gascooke, Alexandra A. Hoops, and Daniel M. Neumar

Single-Batch Production of Recombinant Human Polyclonal Antibodies

We have previously described the development and implementation of a strategy for production of recombinant polyclonal antibodies (rpAb) in single batches employing CHO cells generated by site-specific integration, the SympressTM I technology. The SympressTM I technology is implemented at industrial scale, supporting a phase II clinical development program. Production of recombinant proteins by site-specific integration, which is based on incorporation of a single copy of the gene of interest, makes the SympressTM I technology best suited to support niche indications. To improve titers while maintaining a cost-efficient, highly reproducible single-batch manufacturing mode, we have evaluated a number of different approaches. The most successful results were obtained using random integration in a new producer cell termed ECHO, a CHO DG44 cell derivative engineered for improved productivity at Symphogen. This new expression process is termed the SympressTM II technology. Here we describe proof-of-principle data demonstrating the feasibility of the SympressTM II technology for single-batch rpAb manufacturing using two model systems each composed of six target-specific antibodies. The compositional stability and the batch-to-batch reproducibility of rpAb produced by the ECHO cells were at least as good as observed previously using site-specific integration technology. Furthermore, the new process had a significant titer increase

Inhibition of phosphodiesterase-4 promotes oligodendrocyte precursor cell differentiation and enhances CNS remyelination

The increasing effectiveness of new disease-modifying drugs that suppress disease activity in multiple sclerosis has opened up opportunities for regenerative medicines that enhance remyelination and potentially slow disease progression. Although several new targets for therapeutic enhancement of remyelination have emerged, few lend themselves readily to conventional drug development. Here, we used transcription profiling to identify mitogen-activated protein kinase (Mapk) signalling as an important regulator involved in the differentiation of oligodendrocyte progenitor cells (OPCs) into oligodendrocytes. We show in tissue culture that activation of Mapk signalling by elevation of intracellular levels of cyclic adenosine monophosphate (cAMP) using administration of either dibutyryl-cAMP or inhibitors of the cAMP-hydrolysing enzyme phosphodiesterase-4 (Pde4) enhances OPC differentiation. Finally, we demonstrate that systemic delivery of a Pde4 inhibitor leads to enhanced differentiation of OPCs within focal areas of toxin-induced demyelination and a consequent acceleration of remyelination. These data reveal a novel approach to therapeutic enhancement of remyelination amenable to pharmacological intervention and hence with significant potential for translation