980 resultados para machinery
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Systems which employ underwater acoustic energy for observation or communication are called sonar systems. The active and passive sonars are the two types of systems used for the detection and localisation of targets in underwater. Active sonar involves the transmission of an acoustic signal which, when reflected from a target, provides the sonar receiver with a basis for the detection and estimation. Passive sonar bases its detection and estimation on sounds which emanate from the target itself--Machinery noise, flow noise, transmission from its own active sonar etc.Electroacoustic transducers are used in sonar systems for the transmission and detection of acoustic energy. The transducer which is used for the transmission of acoustic energy is called projector and the one used for reception is called hydrophone. Since a single transducer is not sufficient enough for long range and directional transmission, a properly distributed array of transducers are to be used [9-11].The need and requirement for spatial processing to generate the most favourable directivity patterns for transducer systems used in underwater applications have already been analysed by several investigators [12-21].The desired directivity pattern can be either generated by the use of suitable focussing techniques or by an array of non-directional sensor elements, whose arrangements, spacing and the mode of excitation provide the required radiation pattern or by the combination of these.While computing that the directivity pattern, it is assumed strength of the elements are unaffected by the the source acoustic pressure at each source. However, in closely packed a r r a y s , the acoustic interaction effects experienced among the elements will modify the behaviour of individual elements and in turn will reduce the acoust ic source leve 1 wi t h respect to the maximum t heoret i cal va 1ue a s well as degrade the beam pa t tern. Th i s ef fect shou 1d be reduced in systems that are intended to generate high acoustic power output and unperturbed beam patterns [2,22-31].The work herein presented includes an approach for designing efficient and well behaved underwater transd~cer arrays, taking into account the acoustic interaction effect experienced among the closely packed multielement arrays.Architectural modifications reducing the interaction effect different radiating apertures.
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This thesis is an attempt to explore the problems faced by Indian Women and to examine the ways in which the human rights of women could be better protected in the light of international movements with special reference to national legislation and judicial decisions.The evolution of human rights from early period to Universal Declaration of Human Rights, 1948 is traced in the first chapter. The second chapter deals with the evolution of human rights in India. The evolution of fundamental rights and directive principles and the role played by the Indian Judiciary in enforcing the human rights enumerated in various international instruments dealing with human rights are also dealt with in this chapter. The rights guaranteed to women under the various international documents have been dealt with in the third chapter.It is noticed that the international documents have had their impact in India leading to creation of machinery for protection of human rights. Organised violations of women's rights such as prostitution, devadasi system, domestic violence, sexual harassment at workplaces, the evil of dowry, female infanticide etc. have been analysed in the light of existing laws and decisional jurisprudence in the fourth chapter. The fifth chapter analyses the decisions and consensus that emerged from the world conferences on women and their impact on the Indian Society and Judiciary. The constitutional provisions and legislative provisions protecting the rights of women have been critically examined in the sixth chapter. Chapter seven deals with various mechanisms evolved to protect the human rights of women. The eighth chapter contains conclusions and suggestions.
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The overall focus of the thesis involves the legal protection for consumers of pharmaceutical products.The work on “Legal Protection for Consumers of Pharmaceutical Products” is undertaken to study the legal framework that is existing for this purpose and the functioning of regulating mechanism that is envisaged under it. The purpose of the study is to analyse how far these measures are effective in adequately protecting various aspects of consumer interest. Methodology adopted for the study is analytical.The present study revealed that the theory of freedom of contract is only an ideal relevant when the parties are assumed to be on equal footing.In a more complicated social and economic society, it ceased to have any relevance. Many countries in the world enacted legislations to protect the consumers of pharmaceutical products.The meaning of ‘consumers of drugs’ provided in the law is inclusive and not exhaustive one. The definition of ‘drug’ as interpreted by the courts is comprehensive enough to take in it not only medicines but also substances. The meaning of the word substances has been widened by the interpretation of the courts so as to include all the things used in treatment.The definition of the word ‘consumer’ has been liberally interpreted by the courts so as to provide protective net to a large section of the public.The studies subsequent to this report also revealed that there is a shortage of essential drugs necessary to cure local diseases like tuberculosis and malaria where as drugs containing vitamins and other combinations which are more profitable for the manufacturers are produced and marketed in abundance.The study of the provisions in this regard revealed that the duty of the drug controlling authorities is confined to scrutinize the data of the clinical test already conducted by the sponsor of the drug.Study of the clinical trial procedure under the U.S. law revealed that there is a continuous supervision over clinical trials and controls are provided on the treatment use of an investigational productStudy of the clinical trial procedure under the U.S. law revealed that there is a continuous supervision over clinical trials and controls are provided on the treatment use of an investigational product.the study of the provisions of the Drugs and Cosmetics Act and the rules framed under it revealed that the law in this regard is comprehensive to protect the consumer provided it is sufficiently supported by adequately equipped enforcement machinery.
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This thesis Entitled Socio-Economic Survey of a Specific village.For the purpose of analysis the thesis is divided into nine chapters. The first chapter introduces the subject of study and explains the significance of the study.It also provides the profile of the village. Chapter two deals with the different aspects of agriculture in the village.Chapter three discusses the problems of industrialisation in the Village. Chapter four is on village administration. It elaborates on the services rendered by government machinery in facilitating the development of agriculture and industry in the village.Chapter five explains the ways and means of marketing of village produce, both industrial and agricultural origin. It also explains the relevance of intra village connections in facilitating marketing.Chapter seven provides information regarding the income and expenditure pattern of the village.Chapter eight is on village social life. It explains the social life of the villagers, including religious
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The present work is focused on the organelle and biochemical responses to heavy metal exposure in the fish Oreochromis mossambicus giving particular importance to the metal detoxifying machinery of the organism. The thesis is an outcome of the effort aimed at developing practicable monitoring techniques to deliver guidelines for biological effect monitoring and the need for specific biochemical methods to detect biological effects of heavy metals that can be interpreted in terms of the health status of the individual organism and eventually alterations in vital processes as growth and reproduction. The efficiency of the metal detoxifying metallothioneins which is an attractive tool for biological monitoring, their role as scavengers of trace metal ions and thus in relieving the biological machinery from their toxicity effects are important themes of this study. Efforts have also been made to test the reliability of the spill over hypothesis of the action of metallothioneins (Winge et a1.,1973) and their use as a biological barometer of heavy metal stress.
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The customary laws of Union Territory of Lakshadweep islands are a challenge for judicial institution as well as administrative machinery. With the peculiarities of socio-legal institutions, Lakshadweep system stands apart from the mainstream of legal systems in India. How far do the charismatic modernisation trends flowing into the Lakshadweep society affect the people already protected by the uncodified laws of the past? Many are the issues at this stage. This study analyses them. It examines the growth, evolution and development of the legal system in the islands vis-a-vis the administrative mechanism imposed by the mainland ethos and culture.
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The study covers theFishing capture technology innovation includes the catching of aquatic animal, using any kind of gear techniques, operated from a vessel. Utilization of fishing techniques varies, depending upon the type of fisheries, and can go from a basic and little hook connected to a line to huge and complex mid water trawls or seines operated by large fishing vessels.The size and autonomy of a fishing vessel is largely determined by its ability to handle, process and store fish in good condition on board, and thus these two characteristics have been greatly influenced by the introduction and utilization of ice and refrigeration machinery. Other technological developments especially hydraulic hauling machinery, fish finding electronics and synthetic twines have also had a major impact on the efficiency and profitability of fishing vessels.A wide variety of fishing gears and practices ranging from small-scale artisanal to advanced mechanised systems are used for fish capture in Kerala. Most important among these fishing gears are trawls, seines, lines, gillnets and entangling nets and traps The modern sector was introduced in 1953 at Neendakara, Shakthikulangara region under the initiative of Indo-Norwegian project (INP). The novel facilities introduced in fishing industry by Indo- Norwegian project accordingly are mechanically operated new boats with new fishing nets. Soon after mechanization, motorization programme gained momentum in Kerala especially in Alleppey, Ernakulam and Kollam districts.
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RNA interference (RNAi) is a recently discovered process, in which double stranded RNA (dsRNA) triggers the homology-dependant degradation of cognate messenger RNA (mRNA). In a search for new components of the RNAi machinery in Dictyostelium, a new gene was identified, which was called helF. HelF is a putative RNA helicase, which shows a high homology to the helicase domain of Dicer, to the helicase domain of Dictyostelium RdRP and to the C. elegans gene drh-1, that codes for a dicer related DExH-box RNA helicase, which is required for RNAi. The aim of the present Ph.D. work was to investigate the role of HelF in PTGS, either induced by RNAi or asRNA. A genomic disruption of the helF gene was performed, which resulted in a distinct mutant morphology in late development. The cellular localization of the protein was elucidated by creating a HelF-GFP fusion protein, which was found to be localized in speckles in the nucleus. The involvement of HelF in the RNAi mechanism was studied. For this purpose, RNAi was induced by transformation of RNAi hairpin constructs against four endogenous genes in wild type and HelF- cells. The silencing efficiency was strongly enhanced in the HelF K.O. strain in comparison with the wild type. One gene, which could not be silenced in the wild type background, was successfully silenced in HelF-. When the helF gene was disrupted in a secondary transformation in a non-silenced strain, the silencing efficiency was strongly improved, a phenomenon named here “retrosilencing”. Transcriptional run-on experiments revealed that the enhanced gene silencing in HelF- was a posttranscriptional event, and that the silencing efficiency depended on the transcription levels of hairpin RNAs. In HelF-, the threshold level of hairpin transcription required for efficient silencing was dramatically lowered. The RNAi-mediated silencing was accompanied by the production of siRNAs; however, their amount did not depend on the level of hairpin transcription. These results indicated that HelF is a natural suppressor of RNAi in Dictyostelium. In contrast, asRNA mediated gene silencing was not enhanced in the HelF K.O, as shown for three tested genes. These results confirmed previous observations (H. Martens and W. Nellen, unpublished) that although similar, RNAi and asRNA mediated gene silencing mechanisms differ in their requirements for specific proteins. In order to characterize the function of the HelF protein on a molecular level and to study its interactions with other RNAi components, in vitro experiments were performed. Besides the DEAH-helicase domain, HelF contains a double-stranded RNA binding domain (dsRBD) at its N-terminus, which showed high similarity to the dsRBD domain of Dicer A from Dictyostelium. The ability of the recombinant dsRBDs from HelF and Dicer A to bind dsRNA was examined and compared. It was shown by gel-shift assays that both HelF-dsRBD and Dicer-dsRBD could bind directly to long dsRNAs. However, HelF-dsRBD bound more efficiently to dsRNA with imperfect matches than to perfect dsRNA. Both dsRBDs bound specifically to a pre-miRNA substrate (pre-let-7). The results suggested that most probably there were two binding sites for the proteins on the pre-miRNA substrate. Moreover, it was shown that HelF-dsRBD and Dicer-dsRBD have siRNA-binding activity. The affinities of the two dsRBDs to the pre-let-7 substrate were also examined by plasmon surface resonance analyses, which revealed a 9-fold higher binding affinity of the Dicer-dsRBD to pre-let-7 compared to that of the HelF-dsRBD. The binding of HelF-dsRBD to the pre-let-7 was impaired in the presence of Mg2+, while the Dicer-dsRBD interaction with pre-let-7 was not influenced by the presence of Mg2+. The results obtained in this thesis can be used to postulate a model for HelF function. In this, HelF acts as a nuclear suppressor of RNAi in wild type cells by recognition and binding of dsRNA substrates. The protein might act as a surveillance system to avoid RNAi initiation by fortuitous dsRNA formation or low abundance of dsRNA trigger. If the protein acts as an RNA helicase, it could unwind fold-back structures in the nucleus and thus lead to decreased RNAi efficiency. A knock-out of HelF would result in initiation of the RNAi pathway even by low levels of dsRNA. The exact molecular function of the protein in the RNAi mechanism still has to be elucidated. RNA interferenz (RNAi) ist ein in jüngster Zeit entdeckter Mechanismus, bei dem doppelsträngige RNA Moleküle (dsRNA) eine Homologie-abhängige Degradation einer verwandten messenger-RNA (mRNA) auslösen. Auf der Suche nach neuen Komponenten der RNAi-Maschinerie in Dictyostelium konnte ein neues Gen (helF) identifiziert werden. HelF ist eine putative RNA-Helikase mit einer hohen Homologie zur Helikasedomäne der bekannten Dicerproteine, der Helikasedomäne der Dictyostelium RdRP und zu dem C. elegans Gen drh-1, welches für eine Dicer-bezogene DExH-box RNA Helikase codiert, die am RNAi-Mechanismus beteiligt ist. Das Ziel dieser Arbeit war es, die Funktion von HelF im Zusammenhang des RNAi oder asRNA induzierten PTGS zu untersuchen. Es wurde eine Unterbrechung des helF-Gens auf genomischer Ebene (K.O.) vorgenommen, was bei den Mutanten zu einer veränderten Morphologie in der späten Entwicklung führte. Die Lokalisation des Proteins in der Zelle konnte mit Hilfe einer GFP-Fusion analysiert werden und kleinen Bereichen innerhalb des Nukleus zugewiesen werden. Im Weiteren wurde der Einfluss von HelF auf den RNAi-Mechanismus untersucht. Zu diesem Zweck wurde RNAi durch Einbringen von RNAi Hairpin-Konstrukten gegen vier endogene Gene im Wiltypstamm und der HelF--Mutante induziert. Im Vergleich zum Wildtypstamm konnte im HelF--Mutantenstamm eine stark erhöhte „Silencing“-Effizienz nachgewiesen werden. Ein Gen, welches nach RNAi Initiation im Wildtypstamm unverändert blieb, konnte im HelF--Mutantenstamm erfolgreich stillgelegt werden. Durch sekundäres Einführen einer Gendisruption im helF-Locus in einen Stamm, in welchem ein Gen nicht stillgelegt werden konnte, wurde die Effizienz des Stilllegens deutlich erhöht. Dieses Phänomen wurde hier erstmals als „Retrosilencing“ beschrieben. Mit Hilfe von transkriptionellen run-on Experimenten konnte belegt werden, dass es sich bei dieser erhöhten Stilllegungseffizienz um ein posttranskriptionelles Ereignis handelte, wobei die Stillegungseffizienz von der Transkriptionsstärke der Hairpin RNAs abhängt. Für die HelF--Mutanten konnte gezeigt werden, dass der Schwellenwert zum Auslösen eines effizienten Stillegens dramatisch abgesenkt war. Obwohl die RNAi-vermittelte Genstilllegung immer mit der Produktion von siRNAs einhergeht, war die Menge der siRNAs nicht abhängig von dem Expressionsniveau des Hairpin-Konstruktes. Diese Ergebnisse legen nahe, dass es sich bei der HelF um einen natürlichen Suppressor des RNAi-Mechanismus in Dictyostelium handelt. Im Gegensatz hierzu war die as-vermittelte Stilllegung von drei untersuchten Genen im HelF-K.O. im Vergleich zum Wildyp unverändert. Diese Ergebnisse bestätigten frühere Beobachtungen (H. Martens und W. Nellen, unveröffentlicht), wonach die Mechanismen für RNAi und asRNA-vermittelte Genstilllegung unterschiedliche spezifische Proteine benötigen. Um die Funktion des HelF-Proteins auf der molekularen Ebene genauer zu charakterisieren und die Interaktion mit anderen RNAi-Komponenten zu untersuchen, wurden in vitro Versuche durchgeführt. Das HelF-Protein enthält, neben der DEAH-Helikase-Domäne eine N-terminale Doppelstrang RNA bindende Domäne (dsRBD) mit einer hohen Ähnlichkeit zu der dsRBD des Dicer A aus Dictyostelium. Die dsRNA-Bindungsaktivität der beiden dsRBDs aus HelF und Dicer A wurde analysiert und verglichen. Es konnte mithilfe von Gel-Retardationsanalysen gezeigt werden, dass sowohl HelF-dsRBD als auch Dicer-dsRBD direkt an lange dsRNAs binden können. Hierbei zeigte sich, dass die HelF-dsRBD eine höhere Affinität zu einem imperfekten RNA-Doppelstrang besitzt, als zu einer perfekt gepaarten dsRNA. Für beide dsRBDs konnte eine spezifische Bindung an ein pre-miRNA Substrat nachgewiesen werden (pre-let-7). Dieses Ergebnis legt nah, dass es zwei Bindestellen für die Proteine auf dem pre-miRNA Substrat gibt. Überdies hinaus konnte gezeigt werden, dass die dsRBDs beider Proteine eine siRNA bindende Aktivität besitzen. Die Affinität beider dsRBDs an das pre-let-7 Substrat wurde weiterhin mit Hilfe der Plasmon Oberflächen Resonanz untersucht. Hierbei konnte eine 9-fach höhere Bindeaffinität der Dicer-dsRBD im Vergleich zur HelF-dsRBD nachgewiesen werden. Während die Bindung der HelF-dsRBD an das pre-let-7 durch die Anwesenheit von Mg2+ beeinträchtigt war, zeigte sich kein Einfluß von Mg2+ auf das Bindeverhalten der Dicer-dsRBD. Mit Hilfe der in dieser Arbeit gewonnen Ergebnisse lässt sich ein Model für die Funktion von HelF postulieren. In diesem Model wirkt HelF durch Erkennen und Binden von dsRNA Substraten als Suppressor von der RNAi im Kern. Das Protein kann als Überwachungsystem gegen eine irrtümliche Auslösung von RNAi wirken, die durch zufällige dsRNA Faltungen oder eine zu geringe Häufigkeit der siRNAs hervorgerufen sein könnte. Falls das Protein eine Helikase-Aktivität besitzt, könnte es rückgefaltete RNA Strukturen im Kern auflösen, was sich in einer verringerten RNAi-Effizienz wiederspiegelt. Durch Ausschalten des helF-Gens würde nach diesem Modell eine erfolgreiche Auslösung von RNAi schon bei sehr geringer Mengen an dsRNA möglich werden. Das Modell erlaubt, die exakte molekulare Funktion des HelF-Proteins im RNAi-Mechanismus weiter zu untersuchen.
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Control of protein synthesis is a key step in the regulation of gene expression during apoptosis and the heat shock response. Under such conditions, cap-dependent translation is impaired and Internal Ribosome Entry Site (IRES)-dependent translation plays a major role in mammalian cells. Although the role of IRES-dependent translation during apoptosis has been mainly studied in mammals, its role in the translation of Drosophila apoptotic genes has not been yet studied. The observation that the Drosophila mutant embryos for the cap-binding protein, the eukaryotic initiation factor eIF4E, exhibits increased apoptosis in correlation with up-regulated proapoptotic gene reaper (rpr) transcription constitutes the first evidence for the existence of a cap-independent mechanism for the translation of Drosophila proapoptotic genes. The mechanism of translation of rpr and other proapoptotic genes was investigated in this work. We found that the 5 UTR of rpr mRNA drives translation in an IRES-dependent manner. It promotes the translation of reporter RNAs in vitro either in the absence of cap, in the presence of cap competitors, or in extracts derived from heat shocked and eIF4E mutant embryos and in vivo in cells transfected with reporters bearing a non functional cap structure, indicating that cap recognition is not required in rpr mRNA for translation. We also show that rpr mRNA 5 UTR exhibits a high degree of similarity with that of Drosophila heat shock protein 70 mRNA (hsp70), an antagonist of apoptosis, and that both are able to conduct IRES-mediated translation. The proapoptotic genes head involution defective (hid) and grim, but not sickle, also display IRES activity. Studies of mRNA association to polysomes in embryos indicate that both rpr, hsp70, hid and grim endogenous mRNAs are recruited to polysomes in embryos in which apoptosis or thermal stress was induced. We conclude that hsp70 and, on the other hand, rpr, hid and grim which are antagonizing factors during apoptosis, use a similar mechanism for protein synthesis. The outcome for the cell would thus depend on which protein is translated under a given stress condition. Factors involved in the differential translation driven by these IRES could play an important role. For this purpose, we undertook the identification of the ribonucleoprotein (RNP) complexes assembled onto the 5 UTR of rpr mRNA. We established a tobramycin-affinity-selection protocol that allows the purification of specific RNP that can be further analyzed by mass spectrometry. Several RNA binding proteins were identified as part of the rpr 5 UTR RNP complex, some of which have been related to IRES activity. The involvement of one of them, the La antigen, in the translation of rpr mRNA, was established by RNA-crosslinking experiments using recombinant protein and rpr 5 UTR and by the analysis of the translation efficiency of reporter mRNAs in Drosophila cells after knock down of the endogenous La by RNAi experiments. Several uncharacterized proteins were also identified, suggesting that they might play a role during translation, during the assembly of the translational machinery or in the priming of the mRNA before ribosome recognition. Our data provide evidence for the involvement of La antigen in the translation of rpr mRNA and set a protocol for purification of tagged-RNA-protein complexes from cytoplasmic extracts. To further understand the mechanisms of translation initiation in Drosophila, we analyzed the role of eIF4B on cap-dependent and cap-independent translation. We showed that eIF4B is mostly involved in cap-, but not IRES-dependent translation as it happens in mammals.
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Mit aktiven Magnetlagern ist es möglich, rotierende Körper durch magnetische Felder berührungsfrei zu lagern. Systembedingt sind bei aktiv magnetgelagerten Maschinen wesentliche Signale ohne zusätzlichen Aufwand an Messtechnik für Diagnoseaufgaben verfügbar. In der Arbeit wird ein Konzept entwickelt, das durch Verwendung der systeminhärenten Signale eine Diagnose magnetgelagerter rotierender Maschinen ermöglicht und somit neben einer kontinuierlichen Anlagenüberwachung eine schnelle Bewertung des Anlagenzustandes gestattet. Fehler können rechtzeitig und ursächlich in Art und Größe erkannt und entsprechende Gegenmaßnahmen eingeleitet werden. Anhand der erfassten Signale geschieht die Gewinnung von Merkmalen mit signal- und modellgestützten Verfahren. Für den Magnetlagerregelkreis erfolgen Untersuchungen zum Einsatz modellgestützter Parameteridentifikationsverfahren, deren Verwendbarkeit wird bei der Diagnose am Regler und Leistungsverstärker nachgewiesen. Unter Nutzung von Simulationsmodellen sowie durch Experimente an Versuchsständen werden die Merkmalsverläufe im normalen Referenzzustand und bei auftretenden Fehlern aufgenommen und die Ergebnisse in einer Wissensbasis abgelegt. Diese dient als Grundlage zur Festlegung von Grenzwerten und Regeln für die Überwachung des Systems und zur Erstellung wissensbasierter Diagnosemodelle. Bei der Überwachung werden die Merkmalsausprägungen auf das Überschreiten von Grenzwerten überprüft, Informationen über erkannte Fehler und Betriebszustände gebildet sowie gegebenenfalls Alarmmeldungen ausgegeben. Sich langsam anbahnende Fehler können durch die Berechnung der Merkmalstrends mit Hilfe der Regressionsanalyse erkannt werden. Über die bisher bei aktiven Magnetlagern übliche Überwachung von Grenzwerten hinaus erfolgt bei der Fehlerdiagnose eine Verknüpfung der extrahierten Merkmale zur Identifizierung und Lokalisierung auftretender Fehler. Die Diagnose geschieht mittels regelbasierter Fuzzy-Logik, dies gestattet die Einbeziehung von linguistischen Aussagen in Form von Expertenwissen sowie die Berücksichtigung von Unbestimmtheiten und ermöglicht damit eine Diagnose komplexer Systeme. Für Aktor-, Sensor- und Reglerfehler im Magnetlagerregelkreis sowie Fehler durch externe Kräfte und Unwuchten werden Diagnosemodelle erstellt und verifiziert. Es erfolgt der Nachweis, dass das entwickelte Diagnosekonzept mit beherrschbarem Rechenaufwand korrekte Diagnoseaussagen liefert. Durch Kaskadierung von Fuzzy-Logik-Modulen wird die Transparenz des Regelwerks gewahrt und die Abarbeitung der Regeln optimiert. Endresultat ist ein neuartiges hybrides Diagnosekonzept, welches signal- und modellgestützte Verfahren der Merkmalsgewinnung mit wissensbasierten Methoden der Fehlerdiagnose kombiniert. Das entwickelte Diagnosekonzept ist für die Anpassung an unterschiedliche Anforderungen und Anwendungen bei rotierenden Maschinen konzipiert.
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Dictyostelium discoideum is a social amoeba that serves as a model system for RNA interference and related mechanisms. Its position between plants and animals enables evolutionary snapshot of mechanisms and protein machinery involved in investigated subjects. MiRNAs are small regulatory RNAs that are evolutionary conserved and present in animals, plants, viruses and some prokaryotes. They have roles in development, cell growth and differentiation, apoptosis and their miss-regulation is associated with many diseases such as cancer, neurodegenerative disorders and diabetes. Recently, through sequencing of DNA libraries miRNAs have been discovered in D. discoideum. In this work, it has been shown that heterologues miRNA let-7 can be expressed and processed in D. discoideum. Expression of let-7 miRNA in social amoeba resulted in a strong developmental phenotype suggesting an overload of the processing/silencing system or/and endogenous targets. The various effects on prel-7 strain have been observed and characterized, serving as a background for postulation of miRNA roles. An artificial miRNA system has been established and imposed to D. discoideum, showing that miRNAs in Dictyostelium could mediate gene expression on the level of mRNA stability and on the posttranscriptional level. Furthermore, presence of translational inhibition as a type of gene control was shown for the first time in this organism. Due to it new structures representing co-localities of miRNA and target mRNA have been detected. Taken together, this work shows functional artificial miRNA system and postulates roles of endogenous small RNA in social amoeba.
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Diese Arbeit behandelt Controlled Traffic Farming (CTF) Anbausysteme, bei denen für alle Arbeitsgänge satellitengesteuert immer dieselben Fahrspuren benutzt werden. Lässt sich mit CTF die Belastung des Bodens verringern und die Effizienz von Direktsaat-Anbausystemen steigern? Neben agronomischen und bodenphysikalischen Parametern wurden Auswirkungen von Lenksystemen und Umsetzungsmöglichkeiten von CTF in die Praxis untersucht. Die Analyse einer CTF-Umsetzung unter europäischen Bedingungen mit der Verwendung von Standardmaschinen zeigte, dass sich CTF-Anbausysteme mit den heute zur Verfügung stehenden Maschinen für Dauergrünland, Mähdruschfrüchte und Mais auf kleiner und grösser strukturierten Flächen relativ einfach mechanisieren lassen. Bei Zuckerrüben und Kartoffeln können Kompromisse notwendig sein. Generell erfordern CTF-Anbausysteme eine sorgfältige Planung und Umsetzung in die Praxis. Im dreijährigen Feldversuch (Winterweizen, Wintergerste, Kunstwiese mit Kleegrasmischung) auf einem Lehmboden wurde CTF-Direktsaat mit konventionell zufällig befahrenen Direktsaat- und Pflugverfahren verglichen. Unter CTF zeigte sich eine Differenzierung der nicht, gering und intensiv befahrenen Varianten. Auf dem vorliegenden kompakten Boden mit 1150 mm Jahresniederschlag waren die Unterschiede zwischen den nicht befahrenen Flächen und den mit niedrigem Kontaktflächendruck befahrenen Flächen eher gering. In den nicht befahrenen Flächen entwickelten Eindringwiderstand und Kohlendioxidgehalt der Bodenluft nach drei Jahren signifikant bessere Werte. Bodendichte und Porosität zeigten hingegen keinen eindeutig interpretierbaren Trend. Aufgrund teils suboptimaler Feldaufgänge liess sich keine generelle agronomische Tendenz ableiten. Die intensive Befahrung der Pflegefahrgassen zeigte allerdings klar negative bodenkundliche und planzenbauliche Auswirkungen. Es bietet sich daher an, vor allem für Pflegearbeiten permanent dieselben Fahrspuren zu nutzen. In der Untersuchung zu den Auswirkungen von Lenksystemen zeigten sich signifikante Vorteile von Lenksystemen in einer Verminderung der Fahrerbelastung und einer höheren Lenkgenauigkeit vor allem bei grossen Arbeitsbreiten ohne Spuranreisser. Die meisten anderen Messparameter waren mit Lenksystem leicht vorteilhafter als ohne, unterschieden sich aber nicht signifikant voneinander. Fahrer und naturräumliche Gegebenheiten wie die Schlagform hatten einen wesentlich grösseren Einfluss. Gesamthaft betrachtet erweitert CTF in Kombination mit weiteren Bodenschutzmass-nahmen die Möglichkeiten, Bodenverdichtungen zu vermeiden, den Bedarf an energieintensiver Bodenlocke-rung zu reduzieren und die Entwicklung einer stabileren Bodenstruktur mit höherer Tragfähigkeit zu fördern. Zusammen mit einer an Kultur und Anbausystem angepassten Saatbettbereitung und den in geraden Reihen einfacher durchführbaren mechanischen Pflegemassnahmen ergeben sich gute Voraussetzungen für die Gestaltung agronomisch leistungsfähiger und ökologisch nachhaltiger Anbausysteme.
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For over 1,000 years, the Balinese have developed a unique system of democratic and sustainable water irrigation. It has shaped the cultural landscapes of Bali and enables local communities to manage the ecology of terraced rice fields at the scale of whole watersheds. The Subak system has made the Balinese the most productive rice growers in Indonesia and ensures a high level of food sovereignty for a dense population on the volcanic island. The Subak system provides a vibrant example of a diverse, ecologically sustainable, economically productive and democratic water management system that is also characterized by its nonreliance on fossil fuel derivatives or heavy machinery. In 2012, UNESCO has recognized five rice terraces and their water temples as World Heritage site and supports its conservation and protection. However, the fragile Subak system is threatened for its complexity and interconnectedness by new agricultural practices and increasing tourism on the island.
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This thesis describes several important advancements in the understanding of the assembly of outer membrane proteins of Gram-negative bacteria like Escherichia coli. A first study was performed to identify binding regions in the trimeric chaperone Skp for outer membrane proteins. Skp is known to facilitate the passage of unfolded outer membrane proteins (OMPs) through the periplasm to the outer membrane (OM). A gene construct named “synthetic chaperone protein (scp)” gene was used to express a fusion protein (Scp) into the cytoplasm of E. coli. The scp gene was used as a template to design mutants of Scp suitable for structural and functional studies using site-directed spectroscopy. Fluorescence resonance energy transfer (FRET) was used to identify distances in Skp-OmpA complexes that separate regions in Scp and in outer membrane protein A (OmpA) from E. coli. For this study, single cysteine (Cys) mutants and single Cys - single tryptophan (Trp) double mutants of Scp were prepared. For FRET experiments, the cysteines were labeled with the tryptophan fluorescence energy acceptor IAEDANS. Single Trp mutants of OmpA were used as fluorescence energy donors. In the second part of this thesis, the function of BamD and the structure of BamD-Scp complexes were examined. BamD is an essential component of the β-barrel assembly machinery (BAM) complex of the OM of Gram-negative bacteria. Fluorescence spectroscopy was used to probe the interactions of BamD with lipid membranes and to investigate the interactions of BamD with possible partner proteins from the periplasm and from the OM. A range of single cysteine (Cys) and single tryptophan (Trp) mutants of BamD were prepared. A very important conclusion from the extensive FRET study is that the essential lipoprotein BamD interacts and binds to the periplasmic chaperone Skp. BamD contains tetratrico peptide repeat (TPR) motifs that are suggested to serve as docking sites for periplasmic chaperones such as Skp.
Resumo:
The demand for biomass for bioenergy has increased rapidly in industrialized countries in the recent years. Biogenic energy carriers are known to reduce CO2 emissions. However, the resource-inefficient production of biomass often caused negative impacts on the environment, e.g. biodiversity losses, nitrate leaching, and erosion. The detrimental effects evolved mainly from annual crops. Therefore, the aim of modern bioenergy cropping systems is to combine yield stability and environmental benefits by the establishment of mixed-cropping systems. A particular emphasis is on perennial crops which are perceived as environmentally superior to annual crops. Agroforestry systems represent such mixed perennial cropping systems and consist of a mix of trees and arable crops or grassland within the same area of land. Agroforestry practices vary across the globe and alley cropping is a type of agroforestry system which is well adapted to the temperate zone, with a high degree of mechanization. Trees are planted in rows and crops are planted in the alleyways, which facilitates their management by machinery. This study was conducted to examine a young alley cropping system of willows and two grassland mixtures for bioenergy provision under temperate climate conditions. The first part of the thesis identified possible competition effects between willows and the two grassland mixtures. Since light seemed to be the factor most affecting the yield performance of the understory in temperate agroforestry systems, a biennial in situ artificial shade experiment was established over a separate clover-grass stand to quantify the effects of shade. Data to possible below- and aboveground interactions among willows and the two grassland mixtures and their effects on productivity, sward composition, and quality were monitored along a tree-grassland interface within the alleys. In the second part, productivity of the alley cropping system was examined on a triennial time frame and compared to separate grassland and willow stands as controls. Three different conversion technologies (combustion of hay, integrated generation of solid fuel and biogas from biomass, whole crop digestion) were applied to grassland biomass as feedstock and analyzed for its energetic potential. The energetic potential of willow wood chips was calculated by applying combustion as conversion technique. Net energy balances of separate grassland stands, agroforestry and pure willow stands evaluated their energy efficiency. Results of the biennial artificial shade experiment showed that severe shade (80 % light reduction) halved grassland productivity on average compared to a non-shaded control. White clover as heliophilous plant responded sensitively to limited radiation and its dry matter contribution in the sward decreased with increasing shade, whereas non-leguminous forbs (mainly segetal species) benefited. Changes in nutritive quality could not be confirmed by this experiment. Through the study on interactions within the alleys of the young agroforestry system it was possible to outline changes of incident light, soil temperature and sward composition of clover-grass along the tree-grassland interface. Nearly no effects of trees on precipitation, soil moisture and understory productivity occurred along the interface during the biennial experiment. Considering the results of the productivity and the net energy yield alley cropping system had lower than pure grassland stands, irrespective of the grassland seed mixture or fertilization, but was higher than that for pure willow stands. The comparison of three different energetic conversion techniques for the grassland biomass showed highest net energy yields for hay combustion, whereas the integrated generation of solid fuel and biogas from biomass (IFBB) and whole crop digestion performed similarly. However, due to the low fuel quality of hay, its direct combustion cannot be recommended as a viable conversion technique, whereas IFBB fuels were of a similar quality to wood chip from willow.