Cell-line Engineering of Chinese Hamster Ovary Cells for Low-temperature Culture

Developments in mammalian cell culture and recombinant technology has allowed for the production of recombinant proteins for use as human therapeutics. Mammalian cell culture is typically operated at the physiological temperature of 37°. However, recent research has shown that the use of low-temperature conditions (30-33°) as a platform for cell-culture results in changes in cell characteristics, such as increased specific productivity and extended periods of cell viability, that can potentially improve the production of recombinant proteins. Furthermore, many recent reports have focused on investigating low-temperature mammalian cell culture of Chinese hamster ovary (CHO) cells, one of the principal cell-lines used in industrial production of recombinant proteins. Exposure to low ambient temperatures exerts an external stress on all living cells, and elicits a cellular response. This cold-stress response has been observed in bacteria, plants and mammals, and is regulated at the gene level. The exact genes and molecular mechanisms involved in the cold-stress response in prokaryotes and plants have been well studied. There are also various reports that detail the modification of cold-stress genes to improve the characteristics of bacteria or plant cells at low temperatures. However, there is very limited information on mammalian cold-stress genes or the related pathways governing the mammalian cold-stress response. This project seeks to investigate and characterise cold-stress genes that are differentially expressed during low-temperature culture of CHO cells, and to relate them to the various changes in cell characteristics observed in low-temperature culture of CHO cells. The gene information can then be used to modify CHO cell-lines for improved performance in the production of recombinant proteins.

La pelota no se mancha: Un estudio histórico sobre el desarrollo y la política en FIFA

Esta investigación se centra en la Fédération Internationale de Football Association (FIFA) como organización política. Intenta responder dos interrogantes primordiales: 1) ¿cómo la FIFA ha constituido el poder que tiene actualmente y, así, hacerse del monopolio indiscutido del fútbol? Y 2) ¿cómo ha cambiado en el tiempo la política interna de FIFA y su vínculo con la política internacional? Para lograr esto, se realiza un estudio histórico, basado principalmente en documentos, que intenta caracterizar y analizar los cambios de la organización en el tiempo. Se enfatizan las últimas dos presidencias de FIFA, de João Havelange y Joseph Blatter, como casos de estudio.

Gene transfection of HEK cells on supermacroporous polyacrylamide monoliths: a comparison of transient and stable recombinant protein expression in perfusion culture

Transient and continuous recombinant protein expression by HEK cells was evaluated in a perfused monolithic bioreactor. Highly porous synthetic cryogel scaffolds (10ml bed volume) were characterised by scanning electron microscopy and tested as cell substrates. Efficient seeding was achieved (94% inoculum retained, with 91-95% viability). Metabolite monitoring indicated continuous cell growth, and endpoint cell density was estimated by genomic DNA quantification to be 5.2x108, 1.1x109 and 3.5x1010 at day 10, 14 and 18. Culture of stably transfected cells allowed continuous production of the Drosophila cytokine Spätzle by the bioreactor at the same rate as in monolayer culture (total 1.2 mg at d18) and this protein was active. In transient transfection experiments more protein was produced per cell compared with monolayer culture. Confocal microscopy confirmed homogenous GFP expression after transient transfection within the bioreactor. Monolithic bioreactors are thus shown to be a flexible and powerful tool for manufacturing recombinant proteins.

Unusual censor readings: G.D.R. science fiction and the Ministry of Culture

Covers the censorship by the Hauptverwaltung Verlage und Buchhandel in the Ministry of Culture of science fiction works in East Germany. These ranged from propaganda pieces supporting the scientific-technological revolution and the progress of socialism, to veiled criticisms of the system. The chapter argues that publishers became dependent on the high sales generated by popular culture, creating a relatively safe space for this genre.

Resources, staff beliefs and organizational culture: factors in the use of information and communication technology for adults with intellectual disabilities

Background  Access to, and the use of, information and communication technology (ICT) is increasingly becoming a vital component of mainstream life. First-order (e.g. time and money) and second-order factors (e.g. beliefs of staff members) affect the use of ICT in different contexts. It is timely to investigate what these factors may be in the context of service provision for adults with intellectual disabilities given the role ICT could play in facilitating communication and access to information and opportunities as suggested in Valuing People. Method  Taking a qualitative approach, nine day service sites within one organization were visited over a period of 6 months to observe ICT-related practice and seek the views of staff members working with adults with intellectual disabilities. All day services were equipped with modern ICT equipment including computers, digital cameras, Internet connections and related peripherals. Results  Staff members reported time, training and budget as significant first-order factors. Organizational culture and beliefs about the suitability of technology for older or less able service users were the striking second-order factors mentioned. Despite similar levels of equipment, support and training, ICT use had developed in very different ways across sites. Conclusion  The provision of ICT equipment and training is not sufficient to ensure their use; the beliefs of staff members and organizational culture of sites play a substantial role in how ICT is used with and by service users. Activity theory provides a useful framework for considering how first- and second-order factors are related. Staff members need to be given clear information about the broader purpose of activities in day services, especially in relation to the lifelong learning agenda, in order to see the relevance and usefulness of ICT resources for all service users.

Sport and re/creation: what skateboarders can teach us about learning

This paper explores the role of digital media and creativity in the processes of learning that occur in groups of urban skateboarders. In particular, it examines how the production and consumption of amateur videos contribute to both skaters’ mastery of the techniques of the sport and their integration into the culture of the sport. The data come from an ethnographic study of skateboarders in Hong Kong, which included in-depth interviews, participant observation and the collection of texts and artifacts like magazines, blog entries and amateur skating videos. Skateboarders use video in a number of ways that significantly impact their learning and integration into their communities. They use it to analyze tricks and techniques, to document the stages of their learning and socialization into the group, to set community standards, to build a sense of belonging with their ‘crews’ and to imagine ‘idealized futures’ for themselves and their communities. Understanding the value and function of such ‘semiotic mediation’ in learning and socialization into sport cultures, I suggest, can contribute to helping physical educators design tasks that integrate training in physical skills with opportunities for students to make meaning around their experiences of sport and physical education.

Comparison of agar gel immunodiffusion test, rapid slide agglutination test, microbiological culture and PCR for the diagnosis of canine brucellosis

The performance of the rapid slide agglutination test, with and without 2-mercaptoethanol (RSAT and 2ME-RSAT) and agar gel immunodiffusion test (AGID) was evaluated for the diagnosis of brucellosis in naturally infected dogs. The microbiological culture, PCR and clinical parameters were used as reference. A total of 167 dogs were clinically examined and tested by blood culture, culture of semen/vaginal swab and PCR in blood and semen/vaginal swab. According to the results observed the 167 dogs were divided into three groups: Brucella canis infected dogs (Group 1). B. canis non-infected dogs (Group 2) and dogs with suspected brucellosis (Group 3). The dogs were then tested by RSAT, 2ME-RSAT and AGID. Groups 1 and 2 were used to calculate the diagnostic sensitivity and specificity of the serological tests and the results observed in Group 3 were also discussed. The diagnostic sensitivity of RSAT, 2ME-RSAT and AGID was respectively 70.58%, 31.76%, and 52.94%. The diagnostic specificity of RSAT, 2ME-RSAT and AGID was respectively 83.34%, 100%, and 100%. In dogs with suspected brucellosis 15% were RSAT positive, none was 2ME-RSAT positive and 5% were AGID positive. Although the serological tests are the most commonly used methods for brucellosis diagnosis, a significant proportion of false-negative results were observed highlighting the importance of the direct methods of diagnosis, like blood culture and PCR to improve the diagnosis of canine brucellosis. (c) 2008 Elsevier Ltd. All rights reserved.

Game Audio in Audio Games : Towards a Theory on the Roles and Functions of Sound in Audio Games

For the past few decades, researchers have increased our understanding of how sound functions within various audio–visual media formats. With a different focus in mind, this study aims to identify the roles and functions of sound in relation to the game form Audio Games, in order to explore the potential of sound when acting as an autonomous narrative form. Because this is still a relatively unexplored research field, the main purpose of this study is to help establish a theoretical ground and stimulate further research within the field of audio games. By adopting an interdisciplinary approach to the topic, this research relies on theoretical studies, examinations of audio games and contact with the audio game community. In order to reveal the roles of sound, the gathered data is analyzed according to both a contextual and a functional perspective. The research shows that a distinction between the terms ‘function’ and ‘role’ is important when analyzing sound in digital games. The analysis therefore results in the identification of two analytical levels that help define the functions and roles of an entity within a social context, named the Functional and the Interfunctional levels. In addition to successfully identifying three main roles of sound within audio games—each describing the relationship between sound and the entities game system, player and virtual environment—many other issues are also addressed. Consequently, and in accordance with its purpose, this study provides a broad foundation for further research of sound in both audio games and video games.

Representations of culture in EIL : Cultural representation in Swedish EFL textbooks

The English language has become an international language and is globally used as a lingua franca. Therefore, there has been a shift in English-language education toward teaching English as an interna-tional language (EIL). Teaching from the EIL paradigm means that English is seen as an international language used in communication by people from different linguistic and cultural backgrounds. As the approach to English-language education changes from the traditional native-speaker, target country context, so does the role of culture within English-language teaching. The aim of this thesis is to in-vestigate and analyse cultural representations in two Swedish EFL textbooks used in upper-secondary school to see how they correspond with the EIL paradigm. This is done by focusing on the geograph-ical origin of the cultural content as well as looking at what kinds of culture are represented in the textbooks. A content analysis of the textbooks is conducted, using Kachru’s Concentric Circles of English as the model for the analysis of the geographical origin. Horibe’s model of the three different kinds of culture in EIL is the model used for coding the second part of the analysis. The results of the analysis show that culture of target countries and "Culture as social custom" dominate the cultural content of the textbook. Thus, although there are some indications that the EIL paradigm has influ-enced the textbooks, the traditional approach to culture in language teaching still prevails in the ana-lysed textbooks. Because of the relatively small sample included in the thesis, further studies need to be conducted in order to make conclusions regarding the Swedish context as a whole.

In search of academic identity : physical education, sport science and the field of human movement studies

This thesis addresses the problem of the academic identity of the area traditionally referred to as physical education. The study is a critical examination of the argu ments for the justi cation of this area as an autonomous branch of knowledge. The investigation concentrates on a selected number of arguments. The data collection comprised articles books and proceedings of conferences. The preliminary assessment of these materials resulted in a classi cation of the arguments into three groups. The rst group comprises the arguments in favour of physical education as an academic discipline. The second includes the arguments supporting a science of sport. The third consists of the arguments in favour of to a eld of human movement study. The examination of these arguments produced the following results. (a) The area of physical education does not satisfy the conditions presupposed by the de nition of academic discipline. This is so because the area does not form an integrated system of scienti c theories. (b) The same di culty emerges from the examination of the ar guments for sport science. There is no science of sport because there is no integrated system of scienti c theories related to sport. (c) The arguments in favour of a eld of study yielded more productive results. However di culties arise from the de nition of human movement. The analysis of this concept showed that its limits are not well demarcated. This makes it problematic to take human movement as the focus of a eld of studies. These aspects led to the conclusion that such things as an academic discipline of physical education sport science and eld of human movement studies do not exist. At least there are not such things in the sense of autonomous branches of knowledge. This does not imply that a more integrated inquiry based on several disciplines is not possible and desirable. This would enable someone entering phys ical education to nd a more organised structure of knowledge with some generally accepted problem situations procedures and theories on which to base professional practice.

Brazilian Students and Working Capital: An Analysis of Their Decisions When Using Virtual Market Business Games

This study aims to demonstrate that data from business games can be an important resource for improving efficiency and effectiveness of learning. The proposal presented here was developed from preliminary studies of data from Virtual Market games that pointed the possibility of identifying gaps in learning by analyzing the decisions of students. This proposal helps students to refine their learning processes and equips tutors with strategies for teaching and student assessment. The proposal also complements the group discussion and/or debriefing, which are widely used to enhance learning mediated by games. However, from a management perspective the model has the potential to be erroneous and miss opportunities, which cannot be detected because of the dependence on the characteristics of the individual, such as ability to communicate and work together. To illustrate the proposed technique, data sets from two business games were analyzed with the focus on managing working capital and it was found that students had difficulties managing this task. Similar trends were observed in all categories of students in the study-undergraduate, postgraduate and specialization. This discovery led us to the analysis of data for decisions made in the performance of the games, and it was determined that indicators could be developed that were capable of indentifying inconsistencies in the decisions. It was decided to apply some basic concepts of the finance management, such as management of the operational and non-operational expenditures, as well as production management concepts, such as the use of the production capacity. By analyzing the data from the Virtual Market games using the indicator concept, it was possible to detect the lack of domain knowledge of the students. Therefore, these indicators can be used to analyze the decisions of the players and guide them during the game, increasing their effectiveness and efficiency. As these indicators were developed from specific content, they can also be used to develop teaching materials to support learning. Viewed in this light, the proposal adds new possibilities for using business games in learning. In addition to the intrinsic learning that is achieved through playing the games, they also assist in driving the learning process. This study considers the applications and the methodology used.

Comparison of agar gel immunodiffusion test, rapid slide agglutination test, microbiological culture and PCR for the diagnosis of canine brucellosis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

MINERAL COMPOSITION of RAW MATERIAL, SUBSTRATE and FRUITING BODIES of Pleurotus ostreatus IN CULTURE

In a culture of a Pleurotus ostreatus (oyster mushroom) strain, macro and micronutrients of the raw material and the initial and spent substrates were evaluated. Substrates were formulated with sawdust from Simarouba amara Aubl. and Ochroma piramidale Cav. ex. Lam., crushed Bactris gasipaes Kunth and crushed Saccharum officinarum (sugar cane). Samples were solubilized by means of acid digestion (nitric-peridrol). Ca, Mg, Fe, Cu, Zn and Mn were determined by atomic absorption spectrophotometry, Na and K by atomic emission, and P by colorimetry. The mineral composition of the fruiting body varied with the substrates, which made possible the production of a fruiting body rich in K, P Mg and Fe. Potassium was the mineral with the highest content in the fruiting body in all substrates tested (36.83-42.18g.kg(-1)). There was an increase of protein and mineral content in the spent substrate in relation to the initial one.

Synergistic action of both Aspergillus niger and Burkholderia cepacea in co-culture increases phosphate solubilization in growth medium

Co-inoculation of the fungus Aspergillus niger and the bacterium Burkholderia cepacia was undertaken to understand the interaction between different species of phosphate-solubilizing microorganisms (PSM). PSM were inoculated in a single or mixed (A. nigerB.similar to cepacia) culture. During 9 similar to days of incubation, microbial biomass was enhanced, accompanied with increases in the levels of soluble phosphate and titratable acidity, as well as increased acid phosphatase activity. Production of acids and levels of phosphate solubilization were greater in the co-culture of A.similar to nigerB.similar to cepacia than in the single culture. The quantity of phosphate solubilized by the co-culture ranged from 40.51 +/- 0.60 to 1103.64 +/- 1.21 similar to mu g similar to PO4 3-similar to mL-1 and was 922% higher than single cultures. pH of the medium dropped from 7.0 to 3.0 in the A.similar to niger culture, 3.1 in the co-culture, and 4.2 in the B.similar to cepacia culture. on the third day of postinoculation, acid production by the co-culture (mean 5.40 +/- 0.31 similar to mg NaOH mL-1) was 1990% greater than single cultures. Glucose concentration decreased almost completely (9799% of the starting concentration) by the ninth day of the incubation. These results show remarkable synergism by the co-culture in comparison with single cultures in the solubility of CaHPO4 under in vitro conditions. This synergy between microorganisms can be used in poor available phosphate soils to enhance phosphate solubilization.

Effects of artificial substrate and night-time aeration on the water quality in Macrobrachium amazonicum (Heller 1862) pond culture

The effects of artificial substrate and night-time aeration on the culture of Macrobrachium amazonicum were evaluated in 12 ponds stocked with 45 prawns m-2. A completely randomized design in 2 × 2 factorial scheme with three replicates was used. The combination of factors resulted in four treatments: with substrate and aeration (SA), with substrate and without aeration (SWA), without substrate and with aeration (WSA) and without substrate and aeration (WSWA). The presence of substrate in SA and SWA treatments reduced suspended particles (seston) by ~17.3% and P-orthophosphate by ~50%. The use of aerator (WSA and SA treatments) significantly (P < 0.05) increased the concentration of dissolved oxygen, suspended particles and nutrients in the pond water. These results indicate that the effect of substrate on turbidity and total suspended solids (TSS) values is opposite to the effect of the aerator. The aerators in semi-intensive grow-out M. amazonicum farming lower water quality because they increased the amount of detritus and nutrients in the pond water. On the other hand, the use of artificial substrate reduces turbidity values, chlorophyll a, TSS and P-orthophosphate concentrations. Therefore, the combination of substrate addition and night-time aeration is not interesting because they have opposite effects. © 2013 John Wiley & Sons Ltd.