Följande studie ämnar undersöka hur läromedel i biologi angriper ämnet naturvetenskapens karaktär. Den bygger på de sju aspekter som Norman Lederman använt för att definiera naturvetenskapens karaktär på grund- och gymnasienivå: att det är skillnad mellan observation och slutsats, det är skillnad mellan lagar och teorier, naturvetenskapen är baserad på empiri, den är öppen för förändring, den är kreativ, den är subjektiv och beroende av sitt kulturella sammanhang. Läromedlen analyseras både kvantitativt och med en hermeneutisk brukstextanalys. Resultatet visar att böckerna förmedlar att naturvetenskapen baseras på empiri och till viss del att det är skillnad mellan observationer och slutsatser. De missar dock att förmedla att dagens naturvetenskap är föränderlig, subjektiv och beroende av sitt kulturella sammanhang. Den huvudsakliga slutsatsen är därför att författarna lagt den största vikten vid att förmedla en bild av naturvetenskapens pålitlighet.

System L (SL), a basolateral amino acid transporter, transports large neutral amino acids (LNAAs) in a Na+-independent manner. Previously, we identified two isoforms of transporters: L-type amino acid transporter 1 (LAT1) and 2 (LAT2) and revealed their distinct substrate selectivity and transport properties. In this study, to establish more stable human LAT1 (hLAT1) and LAT2 (hLAT2) in vitro assay systems, we established mouse cell lines stably expressing hLAT1 (S2-LAT1) and hLAT2 (S2-LAT2). Real-time quantitative RT-PCR analysis revealed that S2-LAT1 and S2-LAT2 cells express hLAT1 and hLAT2 mRNAs at 20 - 1000-fold higher levels than those of endogenous mouse Lat1 and Lat2. S2-LAT1 and S2-LAT2 mediated [14C]L-leucine transport properties were measured and corresponded to results observed via Xenopus oocytes. Using these cells, the data demonstrate that hLAT1 and hLAT2 exhibit different characters in the acceptance of α-methyl amino acids and amino acid-related compounds with bulky side chains such as thyroid hormones and melphalan. S2-LAT1 and S2-LAT2 cells are expected to facilitate hLAT1 and hLAT2 substrate recognition research and contribute to drug development by providing an efficient assay system to screen for chemical compounds that interact with hLAT1 and hLAT2.

Virtual Worlds have become an attractive platform for work, play, and learning. Businesses, including the public sector and academia, are increasingly investing their time, money, and attention to understanding the value of virtual worlds as a productivity tool. For example, educators are leading the way with research in Second Life, one of the more popular virtual worlds, as a potentially powerful medium for creating and delivering instruction. Still, little is empirically known about the value of virtual worlds as viable learning platforms. This study examined the instructional potential of Second Life for creating engaging activities, and to investigate the relationship between Second Life and learning in educational settings. It was hypothesized that a positive relationship exists between a learner's level of engagement and achievement. Achievement was assessed as a learner's level of recognition and recall of factual content. It was also hypothesized that a positive relationship exists between a learner's level of engagement and their performance. Performance was assessed as a learner's level of participation, initiative and effort. Additionally, exploratory research was conducted to examine the factors that contributed to both performance and engagement. Lastly, the relationship between other demographic factors of age, Second Life skill level, and ethnicity, with engagement was explored. This research used an empirically tested unit of web-based instructional framework known as a WebQuest. A 3D version, named VWQuest, was created in Second Life. One hundred volunteers completed participation. Using role play, participants participated in a quest for information. While exploring, participants were asked to take photos as evidence of their experiences. Upon completion, they took a knowledge check multiple-choice quiz, and a survey which measured their perceived level of engagement during the activity. Regression analysis indicated no positive correlation between a participant's level of engagement and his or her achievement. However, a positive correlation was found between participants' level of engagement and their performance. Second Life skill level was significantly correlated to performance, and engagement was found to be a mediator between skill level and performance. Most significantly and unexpectedly, participants' performance varied so greatly, the performance rubric was revised four times before it comprehensively captured the diverse range of performances. This evidence suggests that open-ended and creative opportunities to perform yield levels of creativity, engagement, and innovation within immersive platforms, unexpected and far beyond that of traditional instructional settings. Investigating flow dimensions, engagement elements of user control and loss of time were found to be the most significant contributors to performance, and accounted for the greatest amount of variance in explaining performance. Confirmatory factor analysis showed that the flow factors of defined goals and feedback loaded the highest, suggesting a strong relationship between the two factors. Demographic analysis revealed no significant mean difference between gender and engagement, or between age and engagement. The majority of participants were between 40 and 50 and was instructors or educators, not students. For those interested in understanding appropriate and effective instruction in complex, immersive environments, this study brings together new important implications for all of them. Instructional designers may benefit from these findings in their creation of instructional content; instructors may benefit in their curriculum design and teaching methods; and researchers may understand specific facets with instructional potential--engagement factors, technologies, and instructional frameworks--worthy of further investigation.

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Die peroxisomalen ABC-Transporter Pxa1p und Pxa2p sind Halbtransporter. Genetische Studien ergaben Hinweise, dass sie zur Bildung aktiver Transporter heterodimerisieren und am Import von langkettigen Fettsäuren in die Peroxisomen von S. cerevisiae beteiligt sind. Es wurden epitopmarkierte Varianten der Proteine als Komplex isoliert. Damit wurde gezeigt, dass Pxa1p und Pxa2p ein stabiles Heterodimer bilden. Zur Charakterisierung der ATP Bindeeigenschaften wurden die Transporter mit 8-azido-[alpha-32P]-ATP inkubiert und kovalent verknüpft. Dabei konnte gezeigt werden, dass Pxa1p und Pxa2p eine unsymmetrische Bindung des ATP Analogons aufweisen. Pxa2p bindet deutlich mehr azido-ATP als Pxa1p, bei sehr ähnlichen Dissoziationskonstanten. Die reduzierte ATP Bindung von Pxa1p spiegelt sich durch degenerierte Sequenzmotive der an der ATP Bindung beteiligten Sequenzen wieder. Die isolierten ABC-Transporter wurden für ATPase Messungen eingesetzt. Sie zeigten eine basale ATPase Aktivität, die durch Zugabe langkettiger Coenzym A aktivierter Fettsäuren, wie Oleoyl-CoA und Palmitoyl-CoA stimulierbar war. Eine Lysin Mutation im Walker A Motiv von Pxa1p hatte keine Funktionalitätseinbuße zur Folge. Dieselbe Mutation bei Pxa2p führte im Wachstumstest auf Festmedium mit Ölsäure als Kohlenstoffquelle zu einem deutlich verlangsamten Wachstum. Diese Ergebnisse korrespondieren mit der beobachteten unsymmetrischen ATP Bindung von Pxa1p und Pxa2p, da bei dem schwächer bindenden Pxa1p die Mutation wirkungslos blieb. Keine Übereinstimmung war bei den ATPase Aktivitätsmessungen der aufgereinigten Mutanten zu verzeichnen. Beide Mutanten zeigten eine unbeeinträchtigte ATPase Aktivität. Die ABC-Transporter wurden in Proteoliposomen eingebaut und für Transportmessungen mit einem Spin-Label markierten Oleoyl-CoA verwendet. Die Transportmessungen zeigten einen ATP abhängigen Transport, woraus geschlossen wurde, dass Pxa1p-Pxa2p tatsächlich Coenzym A Ester langkettiger Fettsäuren transportiert. / The peroxisomal ABC-transporters Pxa1p and Pxa2p are half transporters. Previous genetic investigations have demonstrated that Pxa1p and Pxa2p have to dimerise in order to build a functional transporter, which is very likely involved in the import of long chain fatty acids into peroxisomes of S. cerevisiae. In this work, tagged versions of the proteins were purified as a complex. This proved for the building of a stable hetero dimer. For characterisation of the ATP binding properties, the transporters were incubated and cross linked with 8-azido-[alpha-32P]-ATP. This revealed an asymmetric binding of the ATP analogue. Pxa2p binds much more azido-ATP, than Pxa1p, while the dissociation constants are rather similar. The poorer ATP binding of Pxa1p is reflected by degenerated sequence motifs in the nucleotide binding fold. The purified ABC-transporters have been used for ATPase assays. They showed a basal ATPase activity, which could be stimulated by addition of long chain fatty acid CoAs, like oleoyl-CoA and palmitoyl-CoA. Mutants with a lysine mutation in the walker A motive of Pxa1p led to no functional impairment, while the corresponding lysine mutation in Pxa2p led to reduced growth on agar plates with oleic acid as sole carbon source. The result corresponds with the ATP binding properties of Pxa1p. Because of the poorer ATP binding, even in the wild type protein, the mutation was not supposed to have a big influence. No accordance was found in respect to the ATPase measurements of the isolated mutant proteins. Both mutants revealed unaffected ATPase activity. The purified ABC-transporters were reconstituted in proteoliposomes and used for translocation assays of a spin-labelled oleoyl-CoA derivative. The measurements revealed an ATP dependent transport of the oleoyl-CoA analogue. This led to the conclusion, that Pxa1p-Pxa2p is indeed the transporter of long chain acetyl CoA esters, which were transported in an ATP dependent manner.

Made available in DSpace on 2016-06-02T20:21:28Z (GMT). No. of bitstreams: 1 4218.pdf: 11765337 bytes, checksum: cdae8703c8d7792c7037dde48c22c345 (MD5) Previous issue date: 2012-04-04 / Financiadora de Estudos e Projetos / Plant cell wall confers to the cell plant, structural support as well as protection against pathogens and phytophagous. Among the cell wall polysaccharides includes pectic substances, which are composed of partially methyl-esterified galacturonic acid residues linked by α-1,4 glycosidic bonds. These enzymes are often synthesized by phytophatogenic micro-organisms for invasion of host plant or by own plant for modeling plant cell wall. The pectic substances are the major component of middle lamella and are natural degraded by pectinases action. Pectin methylesterase (PME) catalysis removes methyl-ester groups, and the Endo-polygalacturonase (Endo-PG) promoves the randomly hydrolysis reaction of α-1,4 bonds. One of the most importante agricultural pest species of the family Curculionidae (Coleoptera: Curculionidae) is Sphenophorus levis, the sugarcane weevil. The larvae of this insect penetrate into the rhizome and build galleries in the stem, decreasing productivity and causing the death of the plant. Large damages to the crop like that are significant in the costs of products derived from sugarcane. Considering the impact of this pest in the sugarcane crop and the absence of efficient method for control, new strategies for controlling are still necessary. The analysis of the cDNA library of S. levis larvaes shows the presence of one PME and one Endo-PG genes that we called Sl-PME and Sl-EndoPG respectively. Considering the importance of studies of insect pests and the extensively use of theses pectinases in different industry fields, we performed the characterization of genomic sequences coding for S. levis pectinases (Sl-Pectinases). It was also carried out the production and characterization of a Sl-PME and a Sl-EndoPG recombinant, expressed in heterologous system. We also accomplished analysis of gene expression by qRTPCR in different stages of development as well as different tissues, and phylogenetic studies between Sl-Pectinases and other pectinases from different kingdoms. The Sl- Pectinases sequences identified, show more similar to homologous insect genes deposited in the GenBank, especially with Sitophilus oryzae. The phylogenetic analysis indicates that the insect group is more correlated with bacteria group and fungi group respectively to PMEs and EndoPGs sequences. Pectinases genomic sequences revealed two introns for Sl-EndoPG gene with 53 and 166 bp, but no one for Sl-PME gene. Both of Sl-Pectinases Recombinant showed catalytic activity. The recombinant Sl-EndoPG shows optimal activity at pH 5,06 ± 0,27 and 49,74 ± 2,49 oC, but extremely low thermostability. For the polygalacturonic acid no-methylated as substract, the enzyme revealed Km = 3,88 mg.mL-1, Vmax = 21.96 μM.s-1 e Kcat = 3.137 s-1; for the citrus pectin partially methylated as substract, the enzyme presented Km = 4,98 mg.mL-1, Vmax = 17,19 μM.s-1 e Kcat = 2.456 s-1. Results in expression analysis suggest that S. levis pectinases have a digestive enzymes role, actting on the midgut. The present work represents the first pectinases of insect produced in Pichia pastoris heterologous system and characterized as optimal conditions of activity, thermostability and kinetic parameters. / A parede celular vegetal confere à célula vegetal suporte estrutural, proteção contra patógenos e fitófagos. Dentre os polissacarídeos da parede celular vegetal são inclusas as substâncias pécticas, as quais são compostas por resíduos de ácido galacturônico, parcialmente esterificados, ligados em série via ligações glicosídicas α- 1,4. As substâncias pécticas são o maior componente da lamela média e são naturalmente degradadas pela ação enzimática das pectinases. A Pectina metilesterase (PME) catalisa a remoção dos grupos metil-ester, e a Endo- Poligalacturonase (Endo-PG) promove a reação de hidrólise aleatória das ligações α- 1,4. Essas enzimas são comumente sintetizadas por micro-organismos fitopatógenos para invasão ao hospedeiro ou pelas próprias plantas para modelamento da parede celular vegetal. Na agricultura, uma das mais importantes espécies pragas da família Curculionidae (Coleoptera: Curculionidae) é o Sphenophorus levis, o bicudo da canade- açúcar. As larvas deste inseto penetram no rizoma e constroem galerias ao longo do colmo, causando queda na produtividade ou até mesmo a morte da planta. Grandes danos na cultura como esses são significativos nos custos de produtos derivados da cana-de-açúcar. Considerando o impacto dessa praga na cultura e a ausência de eficientes métodos de controle, novas estratégias ainda são necessárias no combate à praga. A análise de uma biblioteca de cDNA de larvas do inseto S. levis mostrou a presença de genes codificantes para uma PME e uma Endo-PG, os quais nomeamos de Sl-PME e Sl-EndoPG respectivamente. Devido a importância nos estudos de insetos pragas e a extensa aplicação das pectinases em diversos campos industriais, foi promovida a caracterização das sequências genômicas codificantes para as pectinases de S. levis (Sl-Pectinases). Também foi realizada a produção e caracterização de uma Sl-PME e uma Sl-EndoPG recombinantes, expressas em sistema heterólogo. Além disso, foram conduzidas análises de expressão gênica por qRT-PCR em diferentes estágios de desenvolvimento e diferentes tecidos; e estudos filogenéticos entre as Sl-Pectinases e outras pectinases de diferentes reinos. As sequências das Sl-Pectinases identificadas apresentaram maior similaridade com genes homólogos de insetos depositados no GenBank, principalmente como o Sitophilus oryzae. A análise filogenética indicou que o grupo dos insetos é mais correlacionado com o grupo das bactérias e com o grupo de fungos, respectivamente para as sequências PMEs e Endo-PGs. As sequências genômicas das pectinases revelaram dois introns para Sl-EndoPG com 53 e 166 pb, mas nenhum para o gene Sl- PME. Ambas as Sl-Pectinases Recombinantes apresentaram atividade catalítica. A Sl- EndoPG recombinante mostrou maior atividade em pH 5,06 ± 0,27 e 49,74 ± 2,49 oC, mas baixa termoestabilidade. Para o substrato ácido poligalacturônico não metilado, a enzima revelou Km = 3,88 mg.mL-1, Vmax = 21.96 μM.s-1 e Kcat = 3.137 s-1, para o substrato pectina de citrus parcialmente metilada, a enzima apresentou Km = 4,98 mg.mL-1, Vmax = 17,19 μM.s-1 e Kcat = 2.456 s-1. Os resultados da análise de expressão sugerem que as pectinases de S. levis são enzimas digestivas atuantes no intestino médio. Este trabalho representa as primeiras pectinases de inseto produzidas em sistema heterólogo de Pichia pastoris e caracterizadas quanto a condições ótimas de atividade, termoestabilidade e parâmetros cinéticos.

L’haplodiploïdie chez les hyménoptères leur confère une meilleure résistance aux effets délétères de la dépression de consanguinité. Cependant, certains hyménoptères ont un déterminisme du sexe particulier qui les rend sensibles à cette dépression. Chez ces espèces, le genre des individus dépend de la complémentarité des allèles à un locus donné, le single-locus Complementary Sex Determination (sl-CSD). Les oeufs non fécondés se développent en mâles haploïdes alors hémizygotes au locus de CSD. En revanche les oeufs fécondés donnent des femelles diploïdes s’ils sont hétérozygotes au locus du CSD mais deviennent des mâles diploïdes s’ils sont homozygotes pour ce même locus. Ces derniers sont d’autant plus fréquents dans les populations consanguines où le taux d’homozygotes est élevé. Or, les mâles diploïdes sont dans la majorité des cas non viables ou stériles. La production de tels mâles est de ce fait coûteuse pour les femelles. La faible viabilité des mâles diploïdes s’apparente alors à de la dépression de consanguinité. Par des approches liées à l’écologie comportementale et à l’écologie chimique le projet de thèse se concentre sur l’étude des coûts individuels générés par la production de mâles diploïdes et à l’évolution des comportements sélectionnés en réponse à ces coûts. Dans un premier temps, je me suis intéressée à la fitness des mâles diploïdes et aux répercussions de leur production sur les femelles de la population. Dans un second temps, j’ai étudié les comportements permettant de réduire les coûts de la production des mâles diploïdes. Les individus apparentés représentent les partenaires sexuels où le risque de produire des fils diploïdes est le plus élevé. De ce fait, les comportements d’évitement de la consanguinité devraient être sélectionnés dans ces populations / Haplodiploidy in hymenopterans offers a great resistance to the deleterious effect of inbreeding depression. However, some hymenopterans have a specific sex determination which drives them to this depression. Among those species, the sex of individuals depends on allele complementary at a particular locus; the single-locus Complementary Sex Determinatation (sl-CSD). Unfertilized eggs give birth to haploid males which are hemizygotes at the CSD locus. Conversely, fertilized eggs develop into females when heterozygote at the CSD locus but become diploid males when homozygote at the same locus. Diploid males are especially frequent in inbred populations with a high inbreeding rate. Yet diploid males are generally unviable or sterile. Production of such males is therefore costly for females. Reduced viability of diploid males is then similar to inbreeding depression. Linked with behavioural ecology and chemical ecology approaches, the PhD project starts by focusing on individual costs due to production of diploid males, and then on behaviours that have evolved in response to these costs. As a first step, the fitness of diploid males and its impact on females of the population have been measured. If they are unviable, their production is akin to female mortality. As a second step, we studied behaviours that reduce the cost of diploid male production. Related individuals are sexual partners that are most likely to father diploid sons. Therefore, behavioural inbreeding avoidance may be selected in such populations

Im Sinne einer nachhaltigen und ganzheitlichen Firmenentwicklung lässt sich in der Unternehmenslandschaft ein Trend in Richtung einer kontinuierlichen Erweiterung des unternehmerischen Zielsystems beobachten. Neben etablierten Aspekten, wie dem Qualitätsmanagement und der Arbeitssicherheit, erlangen weitere Themen, wie beispielsweise das betriebliche Umwelt- und Energiemanagement, zunehmend an Bedeutung. Um die Berücksichtigung dieser Aspekte nachzuweisen, steht Unternehmen die Möglichkeit zur Zertifizierung nach funktionsspezifischen Managementsystemnormen offen. Die Einführung und Dokumentation entsprechender Managementsysteme erweist sich jedoch als ein zeitintensiver Prozess, dessen Schwierigkeit vor allem in der unternehmensspezifischen Interpretation der allgemein formulierten Normanforderungen liegt. Strebt ein Unternehmen die Zertifizierung nach mehreren Managementsystemnormen an, so eröffnet sich die Möglichkeit zur Integration der Managementsysteme. Allerdings sind auch die Aufgaben der Integration durch verschiedene Herausforderungen gekennzeichnet, die den Bedarf nach einer systematischen Unterstützung verdeutlichen. Im Bereich der Wirtschaftsinformatik haben sich konzeptuelle Modelle als Instrument zur zielorientierten und verständlichen Beschreibung komplexer Informationssysteme etabliert. Entsprechende Modelle können die Durchdringung und Kommunikation komplexer Sachverhalte durch eine zweckmäßige Abstraktion und Strukturierung vereinfachen und eine Überführung in Anwendungssoftware vorbereiten. Für die vorliegende Dissertation leitet sich unter Berücksichtigung dieser Aspekte folgende zentrale Forschungsfrage ab: Wie kann die konzeptuelle Modellierung bei den Aufgaben der Dokumentation und Integration standardisierter Managementsysteme unterstützen? Der Forschungsmethode des Design Science Research folgend, präsentiert die vorliegende Arbeit funktionsspezifische und funktionsunspezifische Artefakte, die bei der Dokumentation standardisierter Managementsysteme und deren Integration modellbasiert unterstützen. Die Anwendbarkeit der Artefakte wird anhand eines realen Anwendungsfalls demonstriert. Die Ausführungen der vorliegenden Arbeit basieren auf einer Analyse der Anforderungen nach ISO 9001 für Qualitätsmanagementsysteme, ISO 14001 für Umweltmanagementsysteme, ISO 50001 für Energiemanagementsysteme, OHSAS 18001 für Arbeitsschutz- und Arbeitssicherheitsmanagementsysteme sowie des durch die Internationale Organisation für Standardisierung veröffentlichten Annex SL.

Three groups of streams were selected in the area of the Bohemian Massif: the Berounka river with its source streams; the Vltava river with its tributaries the Malše and the Lužnice; the Labe river with its tributaries the Cidlina, the Bystřice and the Javorka. The lithological and tectonical influences to several parameters of the streams were studied. These parameters include: stream gradient, orientation of the stream and sinuosity of the stream. Data were obtained from digital elevation models, aerial photographs, topographical and geological maps. The longitudinal profiles of the streams (in the combination with geological cross-sections), the SL indexes and the gradient/sinuosity graphs were created on the basis of the obtained data. These tools were used for evaluation of the influence of lithology and the tectonics to the streams. The evolution of the streams and the stream-groups was evaluated and compared as well. A strong influence of lithology on the stream gradient was found on most of the analysed streams. The tectonic situation, on the contrary, had strong influence on the orientation of the streams, especially in the resistant rocks. Also, there were selected areas, where was a higher probability of quaternary vertical movements influencing the streams (mountainous areas of the Novohradské...