O objetivo do presente estudo é descrever as características físicas, químicas, morfológicas e funcionais do sêmen do tamanduá-bandeira (Myrmecophaga tridactyla) em cativeiro. Estes resultados representam um dos primeiros requisitos para o conhecimento das características seminais da espécie, visando, em médio prazo, a aplicação de técnicas de reprodução assistida e a formação de bancos de germoplasma. Treze animais foram submetidos à colheita de sêmen em zoológicos no estado de São Paulo Brasil. O método para colheita de sêmen foi a eletroejaculação, semelhante ao realizado em várias espécies silvestres e domésticas, resultando em onze amostras colhidas de nove indivíduos (69,23%). Os testes convencionais indicaram um volume médio de 1,30 ± 0,58 ml, a motilidade média de 33,18 ± 20,14%, o vigor médio de 2,30 ± 0,65, a concentração média de 129,41 ±114,3 x 106 / cm3, o pH médio de 7,44 ± 0,60. A avaliação morfológica apresentou média de defeitos totais de 64,53 ± 19,97%, defeitos maiores 53,77 ± 33,22% e defeitos menores 10,75 ± 6,23%. Nos testes funcionais, a avaliação da integridade da membrana acrossomal mostrou uma média de 16,3 ± 9,3% de danificadas. A avaliação da integridade da membrana plasmática mostrou uma média de 18,9 ± 12,2% de danificadas. A avaliação da atividade mitocondrial revelou as seguintes médias: Classe I = 66,4 ± 19,0%; Classe II = 18,7 ± 9,1%; Classe III = 8,0 ± 6,3%; Classe IV = 3,9 ± 3,1% e Classe V = 3,0 ± 2,9%. A avaliação da fragmentação de cromatina resultou em uma média de 13,21 ± 9,13%. Conclusão: foi demonstrada a aplicabilidade da eletroejaculação na colheita de sêmen de tamanduás-bandeira. Os testes possibilitaram uma análise detalhada das características seminais da espécie, aspectos estes ainda inéditos na literatura mundial. Houve grande variação nos resultados obtidos, refletindo diferenças das condições de manejo e de idade entre os indivíduos e, possivelmente um baixo número amostral estudado. Novos estudos devem ser realizados em animais de vida livre. / The purpose of the present study was to describe the physical, chemical, morphological, and functional features of semen collected from Giant Anteaters (Myrmecophaga tridactyla) raised in captivitya first step toward the feasibility of assisted reproduction techniques and germplasm banks for this species in the mid-term future. Electroejaculation, a method currently applied to a number of wild and domestic species, was performed in 13 animals housed in zoos located in the state of São Paulo, Brazil, yielding 11 samples from 9 animals (69.23%). The following mean values were obtained: volume, 1.30 ± 0.58 mL; motility, 33.18 ± 20.14%; vigor, 2.30 ± 0.65; concentration, 129.41 ± 114.3 x 106 cm-3; pH, 7.44 ± 0.60. Major and minor defects accounted for 53.77 ± 33.22% and 10.75 ± 6.23% of the total, respectively (mean total defect rate, 64.53 ± 19.97%). Damage was found in 16.3 ± 9.3% of acrosomal membranes and 18.9 ± 12.2% of plasma membranes. Mitochondrial activity, categorized as Classes I through V, was 66.4 ± 19.0%, 18.7 ± 9.1%, 8.0 ± 6.3%, 3.9 ± 3.1%, and 3.0 ± 2.9%, respectively. Chromatin fragmentation rate was 13.21 ± 9.13%. Conclusion: The study demonstrated the feasibility of applying electroejaculation to Giant Anteaters for semen collection, allowing for a detailed description of semen features previously unreported for this species. The large variation found in the results reflects differences in management conditions, age range, and possibly the small number of sampled animals, thus warranting further studies with free-ranging specimens.

PURPOSE: To determine, under flow conditions, whether reduced connections between Schlemm’s canal (SC) inner wall (IW) and juxtacanalicular tissue (JCT) cells play a role in giant vacuole (GV) formation; and whether decreased amount of cell margin overlap between adjacent IW cells promotes paracellular pore formation using serial block-face scanning electron microscopy (SBF-SEM). METHODS: Normal human eyes were immersion-fixed (0 mmHg, N=2) or perfusion-fixed (15 mmHg, N=1). Frontal and radial sections of SC were processed for SBF-SEM. IW and JCT cells, GVs, and pores were 3D-reconstructed. In each IW cell, total number of connections with underlying JCT cells/matrix was determined. Total cell margin length (TCML) and zero-overlap length (ZL) of each IW cell were measured to calculate percent zero-overlap length (PZL=ZL/TCML). All data were compared between the eyes fixed at 0 and 15 mmHg. RESULTS: Total number of IW/JCT connections in individual IWs significantly decreased in the eye fixed at 15 mmHg (33±5, N=5 cells) compared to those fixed at 0 mmHg (189±12, N=4 cells, p<0.001). The summed GV volume in individual cells significantly increased in the eye fixed at 15mmHg (218.03±19.65 μm3) compared to those fixed at 0 mmHg (82.33±27.22 μm3, p=0.0043). PZL increased 26.68% (p=0.001) in the eye fixed at 15mmHg vs. those fixed at 0mmHg, and all paracellular pores were found only in regions where the overlap length was 0 μm. CONCLUSIONS: Cellular connections between IW/JCT and IW/IW cells play a role in GV and pore formation in normal human eyes under flow conditions. Our results provide a baseline for future comparison with primary open angle glaucoma eyes.

Marseilleviridae é uma família de vírus gigantes cujos membros infectam amebas de vida livre. Esses vírus têm sido encontrados em amostras ambientais de água, larvas de inseto, torres de resfriamento e mais recentemente em amostras humanas. Eles possuem capsídeo icosaédrico medindo entre 190-250 nm e genoma de DNA dupla fita circular ou linear. Sua replicação ocorre no citoplasma amebiano onde observam-se fábricas virais. Este trabalho tem como objetivo investigar a presença de vírus gigantes em mexilhões-dourados (Limnoperna fortunei) que habitam o Lago Guaíba, Porto Alegre, Brasil. Quarenta indivíduos foram coletados e agrupados em pools de 5 amostras (água interna e corpo, totalizando dezesseis pools). Os pools foram homogeneizados em tampão fosfato, centrifugados e o sobrenadante filtrado em membrana de 0,45μM. Foram cultivadas amebas da espécie Acanthamoeba polyphaga em meio PYG em microplacas de 24 poços, inoculadas com os sobrenadantes, incubadas a 30ºC e examinadas diariamente em busca de efeito citopático (ECP) até 72 horas após a inoculação. Quando CPE era evidente, os sobrenadantes foram coletados e ultracentrifugados através de um gradiente de sacarose de 25%. Um dos dezesseis pools induzindo CPE claro foi submetido à extração de DNA e sequenciamento do genoma viral completo um sequenciador de nova geração (Illumina MiSeq). O genoma do vírus chamado Golden mussel marseillevirus consiste de uma única molécula de DNA de 360610 pb, com um teor de G+C de 43,1%. A análise da sequência de nucleotídeos traduzida revelou a presença de proteínas virais que apresentam homologia com proteínas de outros membros da família Marseilleviridae, como Lausanevirus e vírus Insectomime, porém grande parte do seu genoma não apresenta identidade com proteínas depositadas no banco de dados. A análise filogenética do gene D6/D11 Helicase sugere que este vírus faça parte de uma nova linhagem de marseillevirus. Este é o primeiro estudo que demonstra o isolamento de um vírus gigante a partir de tecidos de mexilhão-dourado e infere que estes vírus estão distribuídos amplamente no meio ambiente. / Marseilleviridae is a family of giant viruses whose members infect free living amoebae. These viruses have been found in environmental samples of water, insect larvae, cooling towers and, more recently, in human samples. They have icosahedral capsids measuring between 190-250 nm and their genome is double-stranded circular or linear DNA. Replication occurs in the host cell cytoplasm, inside the viral factories. This study aims to investigate the presence of giant viruses in tissues of golden mussels (Limnoperna fortunei) that inhabit the Guaiba Lake, Porto Alegre, Brazil. Forty specimens were pooled in groups of 5 specimens (internal water and body, totalizing sixteen pools). The pools were homogenized with phosphate buffer, centrifuged and the supernatant was filtered in 0,45μM. Monolayers of Acanthamoeba polyphaga were cultivated with PYG medium in 24-well microplates, inoculated with the pooled samples, incubated at 30 ºC and examined daily in search for cytopathic effect (CPE) up to 72 hours after inoculation. When CPE was evident, the supernatants were collected, clarified and ultra centrifuged through a 25% sucrose cushion. One out of the sixteen CPE positive pools was submitted to DNA extraction and complete sequencing of the viral genome in a NGS apparatus (Illumina MiSeq). The genome of the virus named Golden mussel marseillevirus consists of a single DNA molecule of 360,610 bp, with a G+C content of 43.1%. The analysis of the translated nucleotide sequence reveals the presence of proteins which are homologs to proteins predicted in other members of the family Marseilleviridae like, e.g. Lausannevirus and Isectomime virus. However, part of the viral genome has no identity with the nucleotide sequences available at the database. The phylogenetic analysis of the D6/D11 Helicase gene suggests that this virus is part of a new lineage of marseillevirus. This is the first study where the isolation of a giant virus from golden mussel tissues is achieved, suggesting that these viruses are widely distributed in the environment.

Cool giant and supergiant stars are among the largest and most luminous stars in the Universe and, therefore, dominate the integrated light of their host galaxies. These stars were extensively studied during last few decades, however their relevant properties like photometric variability and mass loss are still poorly constrained. Understanding of these properties is crucial in the context of a broad range of astrophysical questions including chemical enrichment of the Universe, supernova progenitors, and the extragalactic distance scale. Atmospheres of giant and supergiant stars are characterized by complex dynamics due to different interacting processes, such as convection, pulsation, formation of molecules and dust, and the development of mass loss. Current 1D/3D dynamical model atmospeheres are able to simulate these processes and produce a good agreement with the observed spectral features of evolved stars. However, the models lack constraints and need to be confronted to observables. Dynamical processes in stellar atmospheres impact the formation of spectral lines producing their asymmetries and Doppler shifts. Thus, by studying the line-profile variations on spatial and temporal scales it is possible to reconstruct atmospheric motions in evolved stars. As will be shown in this thesis, a tomographic method is an ideal technique for this purpose. The tomographic method is based on construction and cross-correlation of spectral templates (masks) with observed or synthetic stellar spectra in order to recover velocity fields at different optical depths in the stellar atmosphere.The first part of the thesis further improves the original implementation of the tomographic method. This improvement involves the computation of the contribution function in order to correctly determine an optical depth of formation of spectral lines. The tomographic method is, then, fully validated by applying it to a stellar convection simulation of a red supergiant star and correctly recovering its velocity field throughout the atmosphere. The second part of the thesis applies the tomographic method to the red supergiant star μ Cep in order to constrain its atmospheric motions and relate them to photometric variability. A phase lag (hysteresis) between the effective temperature and the radial velocity variations is revealed with timescales of a few hundred days, similar to photometric ones. A comparison to a stellar convection simulation of a red supergiant star indicates that hysteresis loops are linked to the stochastic shocks generated and shaped by the underlying large-scale convection and may be responsible for photometric variations in μ Cep. The third part of the thesis applies the tomographic method to spectro-interferometric observations of the Mira-type star S Ori. The uniform-disk angular diameters measured at wavelengths contributing to the tomographic masks increase with decrease of an optical depth probed by the masks. This validates the capability of the tomographic method to probe distinct geometrical depths in the stellar atmosphere. The last part of the thesis applies the tomograhic method to the Mira-type star RY Cep and compares the results to those obtained for μ Cep in this thesis. The comparison reveals differences in their behavior in the temperature-velocity plane pointing to the posibility to differentiate between Mira-type and red supergiant stars from their spectroscopic signatures. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

by Cheng Yuen Yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves [106]-113). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.iv / Research out puts --- p.v / Abbreviations --- p.vii / List of Figures --- p.viii / List of Tables --- p.xiii / Table of contents --- p.xiv / Chapter Chapter 1 --- Introduction & Hypothesis / Chapter 1.1. --- General Introduction --- p.1 / Chapter 1.2. --- Hypothesis --- p.4 / Chapter 1.3. --- Objectives --- p.4 / Chapter Chapter 2 --- An Overview of Giant Cell Tumour of Bone / Chapter 2.1. --- Introduction --- p.5 / Chapter 2.2. --- Pathobiological features of GCT --- p.6 / Chapter 2.2.1. --- Radiological appearances and clinical classifications of GCT --- p.7 / Chapter 2.2.2. --- Histological characteristics --- p.10 / Chapter 2.2.3. --- Metastatic GCT --- p.13 / Chapter 2.3. --- Histogenesis of GCT --- p.14 / Chapter 2.4. --- Treatment --- p.19 / Chapter 2.5. --- Summary --- p.22 / Chapter Chapter 3 --- Pharmacological aspect of bisphosphonates / Chapter 3.1. --- Introduction --- p.23 / Chapter 3.2. --- Chemical structures of bisphosphonates --- p.28 / Chapter 3.3. --- Mechanisms and actions --- p.28 / Chapter 3.3.1. --- Bisphosphonates induce osteoclast apoptosis --- p.30 / Chapter 3.3.2. --- Bisphosphonates induce cell apoptosis --- p.32 / Chapter 3.3.3. --- Apoptosis --- p.33 / Chapter 3.3.3.1. --- Morphological characteristic of apoptosis --- p.35 / Chapter 3.4. --- Clinical applications of bisphosphonates --- p.36 / Chapter 3.5. --- Bisphosphonates used in this study --- p.38 / Chapter 3.6. --- Summary --- p.43 / Chapter Chapter 4 --- Materials and methods / Chapter 4.1. --- Introduction --- p.44 / Chapter 4.2. --- Primary GCT cell culture and maintenance --- p.46 / Chapter 4.3. --- Drug preparation --- p.46 / Chapter 4.4. --- MTT assay --- p.47 / Chapter 4.5. --- Annexin-V-flous staining assay --- p.48 / Chapter 4.6. --- Haematoxyline and Eosin staining --- p.51 / Chapter 4.7. --- TUNEL assay (Terminal deoxynucleotidyltrasferase - mediated dUTP-biotin nick end labelling) --- p.52 / Chapter 4.8. --- TEM (Transmission Electron Microscopy) --- p.54 / Chapter 4.9. --- Statistical analysis --- p.54 / Chapter Chapter 5 --- Bisphosphonates induce apoptosis in giant cell tumour of bone -in vitro study / Chapter 5.1. --- Introduction --- p.56 / Chapter 5.2. --- Experimental design --- p.57 / Chapter 5.3. --- Results / Chapter 5.3.1. --- Bisphosphonates reduce cell viability of GCT stromal tumour cell --- p.59 / Chapter 5.3.2. --- Bisphosphonates induce morphological changesin GCT primary culture --- p.59 / Chapter 5.3.3. --- Bisphosphonate significantly induce apoptosis in GCT stromal cells in a dose dependent manner --- p.62 / Chapter 5.4. --- Discussions and Summary --- p.68 / Chapter Chapter 6 --- Bisphosphonates induce apoptosis in giant cell tumour of bone -in vivo study / Chapter 6.1. --- Introduction --- p.73 / Chapter 6.2. --- Experiment design --- p.74 / Chapter 6.3. --- Results / Chapter 6.3.1. --- H & E observations / Chapter 6.3.2. --- Pamidronate significantly induce apoptosis in both osteoclast-like giant cells and stromal tumour cells by TUNEL labelling assay --- p.79 / Chapter 6.3.3. --- Pamidronate induced cellular ultrastructural changes of GCT by TEM examination --- p.83 / Chapter 6.3.4. --- Pamidronate reduce the recurrent characteristic of GCT --- p.95 / Chapter 6.4. --- Discussions and Summary --- p.97 / Chapter Chapter 7 --- Summary and Future Study / Chapter 7.1. --- Summary --- p.101 / Chapter 7.2. --- Future directions --- p.103 / Chapter Chapter 8 --- Reference --- p.105 / Chapter Chapter 9 --- Appendix - solution preparation

La plupart des virus sont connus pour leur capacité à causer des maladies symptomatiques chez l’Homme et chez les autres animaux. Certains d’entre eux sont des grands virus à ADN nommés Virus à Grand ADN Nucléo-Cytoplasmique (NCLDV), rapportés comme infectant les cellules eucaryotiques. Au début du XXI ème siècle, quatre familles ont été définies par Iyer et al. comme ayant une origine commune (groupe monophylétique) : Asfarviridae, Phycodnaviridae, Irido-Ascoviridae et Poxviridae.En 2003, la description d’Acanthamoeba polyphaga mimivirus a cassé un paradigme dans le monde des virus. Par leur taille de particule (450nm), par leur longueur de génomes(supérieure à 1Mb) et leur contenu génique, leur découverte a changé la définition traditionnelle des virus (Lwoff). Depuis 2013 et notamment par les isolements successifs de Pandoravirus,Pithovirus et Mollivirus, ces virus ont été décrits comme possédant de nouvelles propriétés.Leur découverte a été rendue possible grâce à la méthode de co-culture utilisant des protistes, notamment des cellules du genre Acanthamoeba. Cette méthode a été de nombreuses fois modulée par différentes équipes. Dans notre cas, nous avons combiné différentes stratégies appliquées à notre co-culture : la co-culture a été couplée à la cytométrie en flux pour détecter la lyse des protistes. De plus, la cytométrie a été utilisée avec un marqueur à ADN dans le but d’identifier de façon putative le virus et de discriminer les différentes populations virales. Enfin,nous sommes capables de séparer ces populations en utilisant un appareil FACS trieur.L’ensemble de ces techniques a permis l’isolement de nouveaux virus. / Most viruses are known for their ability to cause symptomatic diseases in humans andother animals. Some of them are large DNA viruses named Nucleo-cytoplasmic Large DNAviruses (NCLDV), known for infecting eukaryotic cells. At the beginning of the 21st centuryfour families were defined by Iyer et al. as having a common origin (monophyletic group):Asfarviridae, Phycodnaviridae, Irido-Ascoviridae and Poxviridae.In 2003, the description of Acanthamoeba polyphaga Mimivirus broke this paradigmin the virus world. Because of their particles size (450 nm), their genome size (up to 1Mb),and their gene contents, their discovery changed the traditional definition of viruses (Lwoff).Since 2013 and the successive isolations of Pandoravirus, Pithovirus and Mollivirus; theseviruses have been characterized as possessing various novel properties.Their discoveries have been possible thanks to the co-culture method using protistnotably Acanthamoeba genus cells. This method went through multiple improvements and isemployed by different teams in different ways. In our case and in order to enhance thismethod we combined strategies applied in our co-culture. Indeed, this method consists inusing flow cytometry to detect lysis of protist cells (after all steps of co-culture enrichment).In addition, the flow cytometry was used with a DNA marker in order to identity viruses anddiscriminate viral populations. Then, we were able, using a FACS sorter device, to separatedifferent viral populations from our supernatants.Altogether these techniques have permitted the isolation of new viruses.

No description available.

The thesis deals with the evaluation of habitat preferences of the red deer in the Krkonoše National Park. In the context of Central Europe, the Krkonoše Mountains are an important locality in which many rare and protected ecosystems are located. The current management is trying to restore the species composition of the forest, which was abundant in the past and replaced by atypical spruce species. Red deer (Cervus elaphus), which lives in these forests, has no natural enemy in the Giant Mountains, so it is necessary to control the local population and perform selective management for the purpose of forest protection. For proper management planning it is important to know the life strategies of the red deer: the deer's preffered habitat, physical-geographical characteristics that affect its movement and spatio- temporal use of the area. This information can be obtained through GPS-telemetry collar monitoring. The monitoring campaign in the Giant Mountains was carried out between 2014 and 2018 and the resulting data are used in this work. Information on 71 individuals of red deer are processed and subjected to spatio-temporal analysis, which shows that deer behavior is influenced by both daily and annual cycles. Another aspect influencing its movement is the density and intensity of the use of...

La presente investigación busca determinar cuáles fueron los factores que influyeron en el crecimiento de las exportaciones de Calamar Gigante congelado al mercado español durante los años 2012 – 2016. Primer capítulo, iniciamos con la evolución que ha venido desarrollándose la industria pesquera a nivel internacional y posteriormente como se ha desempeñado la industria en el Perú sobre la extracción, comercialización y exportación de Calamar Gigante congelado. Se investiga si: el cambio climático (niño o niña), el tratado de libre comercio con la Unión Europea, la competitividad, el aporte de las instituciones privadas o públicas al sector pesquero y el mercado de destino y su tendencia de consumo contribuyeron a la exportación de Calamar Gigante Congelado. Segundo capítulo, se determina el planteamiento de la investigación (Problemática e hipótesis), metodológica y tipo de investigación. Las entrevistas fueron realizadas a: Exportadores, Gremio, Instituciones Gubernamentales y Trader. Estos fueron escogidos por ser entidades o empresas representativas al sector. Tercer capítulo, se analizará la información brindada por los entrevistados, la cual nos permitirá obtener un panorama real del sector pesquero potero sobre el problema planteado y nos contribuirá al final del estudio. Cuarto capítulo, se procederá a contrastar las opiniones vertidas en cada entrevista y validar la pertinencia de cada factor con respecto al crecimiento de las exportaciones de pota congelada al mercado español en el periodo de tiempo especificado. Por último, en el quinto capítulo como resultado de la investigación, se presentan conclusiones y recomendaciones. / The following research tries to determinate the factors that influenced the growth of frozen giant squid exports to Spain from 2012 to 2016. First chapter, we begin with the evolution of the fishing industry which has been developing internationally and subsequently how is the situation of the industry in Peru about the extraction, commercialization and export of Frozen Giant Squid. It is investigated whether: climate change (el niño), the free trade agreement with the European Union, competitiveness, the contribution of private or public institutions to the fishing sector and the destination market and its consumption trend contributed to the Export of Frozen Giant Squid. Second chapter, the approach of the research (Problematic and hypothesis), methodological and type of research is determined. The interviews were conducted with: Exporters, Guild, Government Institutions and Trader. These were chosen because they are entities or companies that are representative for the fishery sector. Third chapter, the information provided by the interviewees will be analyzed, which will allow us to obtain a real picture of the fishing sector in the area of the problem and contribute to the end of the study. Fourth chapter, we will proceed to contrast the opinions expressed in each interview and validate the relevance of each factor with respect to the growth of exports of frozen squid to the Spanish market in the specified period of time. Finally, in the fifth chapter as a result of the investigation, conclusions and recommendations are presented. / Tesis

Abstract not available