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Cloning and characterization of gonadotropin receptors in the zebrafish, danio rerio.January 2004 (has links)
Kwok Hin-Fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 84-100). / Abstracts in English and Chinese. / Abstract (in English) --- p.i / Abstract (in Chinese) --- p.iv / Acknowledgement --- p.vi / Table of contents --- p.vii / List of figures --- p.xi / List of tables --- p.xiv / Symbols and abbreviations --- p.xv / List of fish names mentioned in the thesis --- p.xviii / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Gonadotropins / Chapter 1.1.1 --- Structure --- p.1 / Chapter 1.1.2 --- Function --- p.4 / Chapter 1.2 --- Gonadotropin receptor / Chapter 1.2.1 --- Structure --- p.5 / Chapter 1.2.2 --- Expression --- p.7 / Chapter 1.2.3 --- Signaling / Chapter 1.2.3.1 --- cAMP-mediated pathway --- p.7 / Chapter 1.2.3.2 --- Phospholipase C-mediated pathway --- p.9 / Chapter 1.2.4 --- Regulation of expression --- p.12 / Chapter 1.2.5 --- Desensitization of receptors / Chapter 1.2.5.1 --- Uncoupling --- p.13 / Chapter 1.2.5.2 --- Internalization --- p.13 / Chapter 1.3 --- Structure of ovarian follicles --- p.14 / Chapter 1.4 --- The project objectives and long-term significance --- p.16 / Chapter Chapter 2 --- Cloning and Characterization of Zebrafish Follicle-stimulating Hormone (FSH) and Luteinizing Hormone (LH) Receptors ´ؤ Evidence for Distinct Functions of FSH and LH in Follicle Development / Chapter 2.1 --- Introduction --- p.19 / Chapter 2.2 --- Materials and Methods / Chapter 2.2.1 --- Animals and chemicals --- p.22 / Chapter 2.2.2 --- Isolation of total RNA --- p.22 / Chapter 2.2.3 --- Cloning of zebrafish FSHR (zfFSHR) and LHR (zfLHR) cDNA fragments from the zebrafish ovary --- p.23 / Chapter 2.2.4 --- Rapid amplification of 5´ةcDNA ends (5'-RACE) and full-length cDNA --- p.24 / Chapter 2.2.5 --- Isolation of ovarian follicles --- p.25 / Chapter 2.2.6 --- Sampling of the ovaries from sexually immature zebrafish --- p.25 / Chapter 2.2.7 --- Reverse transcription-polymerase chain reaction (RT-PCR) --- p.25 / Chapter 2.2.8 --- Construction of expression plasmids --- p.26 / Chapter 2.2.9 --- Transient transfection and reporter gene assay --- p.27 / Chapter 2.2.10 --- Establishment and characterization of stable zfFSHR or zfLHR-expressing cell lines --- p.28 / Chapter 2.3 --- Results / Chapter 2.3.1 --- Cloning of FSHR and LHR cDNA from the zebrafish ovary --- p.29 / Chapter 2.3.2 --- Functional characterization of zfFSHR and zfLHR --- p.30 / Chapter 2.3.3 --- Expression of zfFSHR and zfLHR during sexual maturation --- p.31 / Chapter 2.3.4 --- Stage-dependent expression of zfFSHR and zfLHR in the ovarian follicles --- p.32 / Chapter 2.4 --- Discussion --- p.33 / Chapter Chapter 3 --- Down-regulation of FSHR and LHR Expression in the Zebrafish Follicle Ceils by Gonadotropin (hCG) and Its Sigaling Mechanism / Chapter 3.1 --- Introduction --- p.51 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Animals --- p.54 / Chapter 3.2.2 --- Chemicals and hormones --- p.54 / Chapter 3.2.3 --- Primary follicle cell culture --- p.55 / Chapter 3.2.4 --- Total RNA isolation --- p.55 / Chapter 3.2.5 --- "Validation of semi-quantitative RT-PCR assays for FSHR, LHR and GAPDH" --- p.56 / Chapter 3.2.6 --- Data analysis --- p.57 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Validation of semi-quantitative RT-PCR assays --- p.57 / Chapter 3.3.2 --- Gonadotropin regulation of FSHR and LHR expression in cultured zebrafish ovarian follicle cells --- p.58 / Chapter 3.3.3 --- Effect of db-cAMP and forskolin on FSHR and LHR expression --- p.59 / Chapter 3.3.4 --- Effects of H89 on hCG-induced suppression of FSHR and LHR expression --- p.60 / Chapter 3.4 --- Discussion --- p.60 / Chapter Chapter 4 --- General Discussion --- p.75 / Chapter 4.1 --- Cloning of zebrafish FSHR and LHR cDNAs and demonstration of receptor specificity --- p.77 / Chapter 4.2 --- Evidence for the differential expression of FSHR and LHR in the zebrafish ovarian and follicle development --- p.78 / Chapter 4.3 --- Down-regulation of FSHR and LHR expression in the zebrafish follicle cells by gonadotropin (hCG) --- p.79 / Chapter 4.4 --- Future research direction --- p.80 / References --- p.84
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Cloning and characterization of gonadotropins in the zebrafish, Danio rerio.January 2004 (has links)
So Wai-Kin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 100-127). / Abstracts in English and Chinese. / Acknowledgement --- p.I / Abstract (in English) --- p.II / Abstract (in Chinese) --- p.IV / Table of contents --- p.VI / List of Figures --- p.X / Symbols and Abbreviations --- p.XII / List of fish names mentioned in the thesis --- p.XIV / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Pituitary --- p.1 / Chapter 1.2 --- Gonadotropins --- p.1 / Chapter 1.2.1 --- Structure --- p.2 / Chapter 1.2.2 --- Signaling --- p.3 / Chapter 1.2.3 --- Expression --- p.5 / Chapter 1.2.4 --- Functions --- p.7 / Chapter 1.2.4.1 --- Gonadotropin actions on gametogenesis --- p.7 / Chapter 1.2.4.2 --- Gonadotropin actions on steroidogenesis --- p.8 / Chapter 1.2.5 --- Regulation --- p.9 / Chapter 1.2.5.1 --- Neuroendocrine control --- p.10 / Chapter 1.2.5.1.1 --- Gonadotropin-releasing hormone (GnRH) --- p.10 / Chapter 1.2.5.1.2 --- Dopamine (DA) --- p.12 / Chapter 1.2.5.2 --- Gonadal steroid feedback --- p.12 / Chapter 1.2.5.2.1 --- Positive feedback --- p.13 / Chapter 1.2.5.2.2 --- Negative feedback --- p.14 / Chapter 1.2.5.3 --- Paracrine regulators within pituitary --- p.15 / Chapter 1.3 --- Objectives of the present study --- p.16 / Chapter Chapter 2 --- "Molecular Cloning and Functional Characterization of Zebrafish FSHβ, LHβ and GTHα subunits" / Chapter 2.1 --- Introduction --- p.19 / Chapter 2.2 --- Materials and methods --- p.21 / Chapter 2.2.1 --- Chemicals --- p.21 / Chapter 2.2.2 --- Animals --- p.21 / Chapter 2.2.3 --- Genomic DNA isolation --- p.22 / Chapter 2.2.4 --- Total RNA isolation --- p.22 / Chapter 2.2.5 --- Cloning of zebrafish FSHp,LHβ and GTHa fragments --- p.23 / Chapter 2.2.5.1 --- LHβ and GTHα --- p.23 / Chapter 2.2.5.2 --- FSHβ --- p.23 / Chapter 2.2.6 --- "5'- and 3'-RACE of zebrafish FSHp, LHβ and GTHα subunits" --- p.24 / Chapter 2.2.7 --- Construction of expression constructs --- p.25 / Chapter 2.2.8 --- Cell culture and transfection of Flp-In´ёØ CHO cell --- p.26 / Chapter 2.2.9 --- Recombinant production of zebrafish FSH and LH --- p.27 / Chapter 2.2.10 --- Reverse transcription-polymerase chain reaction (RT-PCR) analysis --- p.27 / Chapter 2.2.11 --- Northern blot hybridization --- p.28 / Chapter 2.2.12 --- SEAP reporter gene assay --- p.28 / Chapter 2.2.13 --- Data analysis --- p.29 / Chapter 2.3 --- Results --- p.30 / Chapter 2.3.1 --- "Cloning of zebrafish FSHβ, LHβ and GTHα subunits" --- p.30 / Chapter 2.3.2 --- "Expression of zebrafish FSHp, LHβ and GTHα in the zebrafish pituitary" --- p.31 / Chapter 2.3.3 --- Recombinant production of zebrafish FSH and LH --- p.32 / Chapter 2.3.4 --- Functional analysis of zebrafish FSH and LH --- p.33 / Chapter 2.4 --- Discussion --- p.34 / Chapter Chapter 3 --- "Spatial Expression Patterns of Zebrafish FSHβ, LHβ and GTHα Subunits in the Pituitary and Their Temporal Expression Profiles during Sexual Maturation and Ovulatory Cycle" / Chapter 3.1 --- Introduction --- p.58 / Chapter 3.2 --- Materials and methods --- p.61 / Chapter 3.2.1 --- Chemicals --- p.61 / Chapter 3.2.2. --- Animals --- p.62 / Chapter 3.2.3 --- Total RNA isolation from zebrafish pituitaries and reverse transcription --- p.62 / Chapter 3.2.4 --- Validation of RT-PCR on single pituitary --- p.63 / Chapter 3.2.5 --- Real-time PCR --- p.64 / Chapter 3.2.6 --- Tissue preparation for in situ hybridization --- p.64 / Chapter 3.2.7 --- In situ hybridization --- p.65 / Chapter 3.2.8 --- Data analysis --- p.66 / Chapter 3.3 --- Results --- p.66 / Chapter 3.3.1 --- "PCR amplification of FSHβ, LHβ and GTHα and GAPDH in single zebrafish pituitary" --- p.67 / Chapter 3.3.2 --- "Establishement of real-time RT-PCR for zebrafish FSHβ, LHβ and GTHa and GAPDH" --- p.67 / Chapter 3.3.3 --- "Temporal expression profiles of zebrafish FSHβ, LHβ and GTHα subunits during sexual maturation" --- p.67 / Chapter 3.3.4 --- "Temporal expression profiles of zebrafish FSHp, LHβ and GTHα subunits during ovulatory cycle" --- p.68 / Chapter 3.3.5 --- "In situ hybridization of zebrafish FSHβ, LHβ and GTHα" --- p.69 / Chapter 3.4 --- Discussion --- p.70 / Chapter Chapter 4 --- General Discussion / Chapter 4.1 --- Cloning of zebrafish gonadotropin subunit cDNAs --- p.91 / Chapter 4.2 --- Bioactivity and receptor specificity of recombinant zebrafish FSH and LH --- p.91 / Chapter 4.3 --- Expression of gonadotropin subunits during zebrafish sexual maturation and ovulatory cycle --- p.92 / Chapter 4.4 --- "Localization of FSHβ, LHβ and GTHα subunits in zebrafish pituitary" --- p.93 / Chapter 4.5 --- Contributions of the present study --- p.94 / Chapter 4.6 --- Future prospects --- p.95 / References --- p.100
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Conquering Taiwan by Mind-PRC Taiwan's Psychology of Political CommunicationTsai, Tzung-je 13 August 2007 (has links)
This research systemizes PRC¡¦s Taiwan policy in the perspective of psychology of political communication. First of all, this research is found that there are four features in psychological warfare conducted by PRC after reviewing the Western, ancient Chinese, and Chinese military classics: justice warfare, army controlled by the politics, the people warfare, and influx on strategies.
The basic levels in mutual fight can be analyzed as following: conducting and controlling relationship, civil-martial relationship, state-society relationship, and foreign relationship. The rival may ¡§disturb¡¨ the relationships by three psychological principles of political communication: the attachment principle, the inducement principle, and the coercion principle. After deciding the principle, there are four kinds of ¡§tools¡¨ available; including Taiwan affairs, mass communication, information based psychological warfare, and military threat. Most important, PRC use the tools in a flexible way.
The psychology of political communication in PRC¡¦s Tawian policy is successful by adopting two stable principles: Emancipation/ Unity/ Anti-independency Principle; One China/ One Country Two Systems Principle. The bottom line of principle can be shifting, but never quit.
From the perspective of psychology of political communication, this Taiwan policy can be focused on the source and the content of information, and the effect of receivers. This model can be concluded as ¡§organism¡¨, there are holism, correlation, multilevel, openness, and dynamics in this model.
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Materialisations of space: phenomenological-archaeological investigations concerning the relations between the human organism, space and technologyWoelert, Peter Christian, History & Philosophy, Faculty of Arts & Social Sciences, UNSW January 2008 (has links)
In my thesis I investigate the impact the conceptual domestication and material technical transformation of space and movement has had on the behaviour of the human organism and the way it relates to its environment. In doing so l will examine the formation and structure of three non-identical yet interrelated forms of human space; rational-geometrical space, lived space, and technologically mediated space, I combine a phenomenological approach, which allows for an analysis of the horizontal relation between embodied organism and environment, with an archaeological perspective that traces the genealogy of specific symbolic and technological formations viewed in their nexus with lived, embodied behaviour. I argue that both the process of the conceptual domestication of space, particulady in the form of what Husserl refers to as the tendency of rationalisation and technisation, as well as the concrete technological transformation of the spatial environment, come into being and develop in a comparable way, While both initially directly or indirectly presuppose the perceptual' and motor activity of the embodied human organism, their subsequent development is tendentiously characterised by a relative departure from the human body in lieu of an extra-somatic organisation and materialisation of sense and behaviour. The implications for the behaviour of the individual human organism are ambivalent. On the one hand, the increasing uncoupling of technology and conceptual systems from human embodiment, has allowed for a rapid development of the human's overall technical and symbolic capacities, The result is an expansive material and symbolic 'humanisation' (Leroi-Gourhan) of the organism's behavioural and geographical environment. On the other hand, the very same process entails a behavioural regression with regard to the human organism's sensorimotor activity. I argue, by way of the former, that this may entail a constriction of the human organism's cognitive and imaginative capacities, potentially threatening its individuality.
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Fresh Face >>> Body 2.0 : Handens minne + Digitala dimensionerWollin, Elin January 2017 (has links)
"My goal with this project is to digital print a Science Fiction inspired print on fabric that provides an optical illusion with lenticular technology. The fabric will be presented with five curtain drapings, two in motion with motorized movable suspensions. The Print design is going to be created with a science fiction/cyberpunk/transhumanism atmosphere. I’m inspired by the relation between body and machine. The textile field is drawn to smart textiles where the idea of function, artificial intelligence and virtual reality is central. There are already textiles that are designed to react to the wearers needs. And soon it will be possible for the online shopper to purchase customized clothes by recreating themselves as an avatar through 3Dscanning. With this as inspiration my digital print is designed. Technically I will produce a fabric using the lenticular technology that creates an optical illusion. In order to produce the fabric, I need to work with the prints in Photoshop and then digitally print them on fabric. So that the fabric could be able to obtain an optical effect, a lens made of silicone will be laminated on top. The silicone lens is cast in a form which is sketched and made as a model in Rhino and then cut out from a metal block with a milling cutter. The composition will be presented as five curtain drapings. Two of them moves with electric winches. They will hang separately but communicate in a spatial constellation. Like organs the draperies spread out and fall back again. Here, as in the print design, the body and the machine will be crucial. The focus is also on the fabric's tactile character, drama and historical heritage."
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Magnetoelectric Nanoparticles: Paradigm Shift in Biomolecular DiagnosticsKhanal, Chooda Mani 08 November 2016 (has links)
Recently discovered multiferroic nanoparticles (MFNs) known as magnetoelectric nanoparticles (MENs) promise to revolutionize next-generation biomedical diagnostic techniques and enable rapid and cost-effective screening of lethal diseases. Due to the quantum-mechanically caused intrinsic magnetoelectric (ME) effect, these nanoparticles display strongly coupled magnetic and electric moments and thus provide a unique pathway to monitor and optionally control intrinsic characteristics of bio organisms via application of external magnetic fields. Due to the ME effect, when placed in the microenvironment of a biomolecular sample, MENs’ magnetic properties change to reflect the molecular nature of the sample. Such an unprecedented intrinsic connection to cells and microorganism’s intrinsic characteristics can pave a way to a new diagnostic paradigm.
A novel concept of portable biomolecular screening device based on continuous wave nuclear magnetic resonance is being studied to identify microorganisms like bacteria, virus and cancer cell lines. For the first time, MENs have been used to modify NMR spectra and thus enable identification of different cancer cell lines from each other as well as from their normal counterparts at the sub-cellular level. This approach can be used for both in vitro and in vivo diagnostics. In addition, blood samples of Pulmonary Arterial Hypertension (PAH) patients with severe heart and lung conditions were used to study the activity of MENs with prothrombin, the clotting factor of the blood. MENs has significant interaction with prothrombin. MENS can be used for diagnostics of several diseases associated with blood and cancer and it may be used as a therapeutic agent. The experiment proved that the MENs are safe and nontoxic carriers of therapeutic drugs.
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Ontology Design Patterns for Combining Pathology and Anatomy: Application to Study Aging and Longevity in Inbred Mouse StrainsAlghamdi, Sarah M. 13 May 2018 (has links)
In biomedical research, ontologies are widely used to represent knowledge as well as to annotate datasets. Many of the existing ontologies cover a single type of phenomena, such as a process, cell type, gene, pathological entity or anatomical structure. Consequently, there is a requirement to use multiple ontologies to fully characterize the observations in the datasets. Although this allows precise annotation of different aspects of a given dataset, it limits our ability to use the ontologies in data analysis, as the ontologies are usually disconnected and their combinations cannot be exploited. Motivated by this, here we present novel ontology design methods for combining pathology and anatomy concepts. To this end, we use a dataset of mouse models which has been characterized through two ontologies: one of them is the mouse pathology ontology (MPATH) covering pathological lesions while the other is the mouse anatomy ontology (MA) covering the anatomical site of the lesions. We propose four novel ontology design patterns for combining these ontologies, and use these patterns to generate four ontologies in a data-driven way.
To evaluate the generated ontologies, we utilize these in ontology-based data analysis, including ontology enrichment analysis and computation of semantic similarity. We demonstrate that there are significant differences between the four ontologies in different analysis approaches. In addition, when using semantic similarity to confirm the hypothesis that genetically identical mice should develop more similar diseases, the generated combined ontologies lead to significantly better analysis results compared to using each ontology individually. Our results reveal that using ontology design patterns to combine different facets characterizing a dataset can improve established analysis methods.
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Engineering A New Form Of Enclosure: International Convergence In Gmo RegulationAltif, Jessica 01 January 2010 (has links)
As society begins to recognize its impact on ecological systems, the belief that modern political institutions can offer a sense of control and certainty, as well as protect the health of its citizens, is increasingly questioned. In an era of uncertainty, faith in science and technology to alleviate industrial impacts on the environment is often embraced by policymakers yet questioned by the public who see the authoritative role of the sciences in the political sphere as contributing to global risk. The development of biotechnology, specifically genetically modified food, places an anthropocentric focus on resolving and/or adapting to environmental degradation, further reflecting an adherence to the dominant social paradigm to address the consequences of modernization. In order to explicate the dualism of human/nature relations inherent in biotechnology, the focus of this research provides an exploration into two competing paradigms of genetically modified organism (GMO) regulatory policy: scientific rationality and social rationality. Through a careful examination of the evolution of GMO regulation in the United States and the European Union, the precarious relationships between science and politics and progress and precaution reveal an actual convergence instead of divergence between these two actors in the international system. Although existing literature proclaims a division between the values and ethics of U.S. and EU environmental policy, the end result of this comparison in GMO regulation illustrates that in both the risk assessment and precautionary approaches, nature is still viewed as an instrument for advancing enclosure of the commons.
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Analysis of the Susceptibility, Prevalence, and Pathogenicity of the Opportunistic Pathogen AcanthamoebaShoff, Megan E. January 2008 (has links)
No description available.
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Effect of Growth Horomone on Rainbow Trout: An Ecological Assessment of the Potential Impact of Genetic Engineering on Organism Design / An Ecological Assessment of Genetic EngineeringJobin, Richard 08 1900 (has links)
Trout genetically engineered to possess extra functioning growth homone genes were simulated, using injections of fish and bovine growth hormone. This was done to test potential and ecological impact of such genetic engineering on trout. Through analysis of growth and dry-mass budgets, it was determined that elevated levels of growth hormone resulted in increased growth and consumption. Potential constraints pertaining to respiration, density and evolutionary history were considered. Surprisingly, bovine growth hormone appeared to act as a super-normal stimuli, being more potent than the natural trout hormone. In fact the natural hormone inhibited growth at high dosages. Juvenile growth rates were less sensitive to elevated levels of growth hormone than those in more mature fish. Thus, improved growth was achieved by altering its normal ontogeny. This suggests that duplicating copies of growth hormone gene in an organism is not equivalent to a speciation event. Evidently, other manipulations would be required to increase the intrinsic metabolic power. Growth rate apparently is used as a cue for determining a particular ontogenetic trajectory (Stearns and Crandall, 1984). The fact that various species conform to one of at least four plastic developmental trajectories that are shaped by natural constraints and mortality patterns, means that growth hormone may elicit different responses in different species. The findings and implications of this study underscore the importance of using the holistic scope of ecology to achieve effective and efficient genetic engineering of target species. / Thesis / Master of Science (MS)
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