Anthropogenic activities are increasing inorganic nitrogen (N) loadings to lakes in the northern hemisphere. In many boreal lakes phytoplankton are N limited, wherefore enhanced N input may affect the productivity of pelagic food webs. Simultaneously, global change causes increased inflows of terrestrial dissolved organic carbon (DOC) to boreal lakes. Between clear and humic lakes, whole lake primary and consumer production naturally differs. However, research is inconclusive as to what controls pelagic production in these lakes. Further, it is unclear how DOC affects the response of the pelagic food web to enhanced inorganic N availability. The overarching goal of this thesis was to study the effects of inorganic N and organic C for pelagic food webs in boreal lakes. In the thesis, I first identified the main drivers of pelagic production during summer in eight non-manipulated Swedish boreal lakes with naturally low or high DOC. Then I investigated how increased N availability affects the pelagic food chain, and how the response differs with DOC. Therefore, whole lake inorganic N fertilization experiments were conducted in six Swedish boreal lakes across a DOC gradient (low, medium, high) divided into three lake pairs (control, N enriched) with one reference and two impact years. In each lake, I also investigated the response of zooplankton growth using in situ mesocosm experiments excluding planktivores. I found that humic boreal lakes had lower phytoplankton production and biomass than clear water lakes. Further, phytoplankton community composition and food quality differed with DOC. However, high DOC did not reduce pelagic energy mobilization or zooplankton biomass, but promoted a higher dominance of cladoceran relative to copepod species. N addition clearly enhanced phytoplankton biomass and production in the experimental lakes. However, this stimulating N effect decreased with DOC as caused by light limitation. Further, the newly available phytoplankton energy derived from N addition was not efficiently transferred to zooplankton, which indicates a mismatch between producer energy supply and consumer energy use. Indeed, the mesocosm experiment revealed that decreased food quality of phytoplankton in response to N addition resulted in reduced food web performance, especially in clearer lakes. In humic lakes, zooplankton production and food web efficiency were clearly more resilient to N addition. In summary, my thesis suggests that any change in the landscape that enhances inorganic N availability will especially affect pelagic food webs in clear water lakes. In contrast, brownification will result in more lakes being resilient to eutrophication caused by enhanced N deposition.

Droplet microfluidics is an emerging technology for analysis of single cells and biomolecules at high throughput. The controlled encapsulation of particles along with the surrounding microenvironment in discrete droplets, which acts as miniaturized reaction vessels, allows millions of particles to be screened in parallel. By utilizing the unit operations developed to generate, manipulate and analyze droplets, this technology platform has been used to miniaturize a wide range of complex biological assays including, but not limited to, directed evolution, rare cell detection, single cell transcriptomics, rare mutation detection and drug screening. The aim of this thesis is to develop droplet microfluidics based methods for analysis of single cells and nucleic acids. In Paper I, a method for time-series analysis of mammalian cells, using automated fluorescence microscopy and image analysis technique is presented. The cell-containing droplets were trapped on-chip and imaged continuously to assess the viability of hundreds of isolated individual cells over time. This method can be used for studying the dynamic behavior of cells. In Paper II, the influence of droplet size on cell division and viability of mammalian cell factories during cultivation in droplets is presented. The ability to achieve continuous cell division in droplets will enable development of mammalian cell factory screening assays in droplets. In Paper III, a workflow for detecting the outcome of droplet PCR assay using fluorescently color-coded beads is presented. This workflow was used to detect the presence of DNA biomarkers associated with poultry pathogens in a sample. The use of color-coded detection beads will help to improve the scalability of the detection panel, to detect multiple targets in a sample. In Paper IV, a novel unit operation for label-free enrichment of particles in droplets using acoustophoresis is presented. This technique will be useful for developing droplet-based assays that require label-free enrichment of cells/particles and removal of droplet content. In general, droplet microfluidics has proven to be a versatile tool for biological analysis. In the years to come, droplet microfluidics could potentially be used to improve clinical diagnostics and bio-based production processes. / <p>QC 20160926</p>

This paper studies the nuclear technology and evaluates the most likely technical developments to come until 2030. It examines the features of Czech and French nuclear programs, compares the structures of nuclear sectors and estimates the advantages & drawbacks of further developments in both countries. The paper assesses if certain developments of the nuclear sector in one country can be used in the other country and vice-versa. Finally, proposes a set of recommendations for both countries regarding the development of their nuclear program.

Thesis deals with the use of mediated learning experience in the focus on the communication skills of preschool children. Thesis follows up the theoretical knowledges of preschool education especially content of it and related to main documentes. Thesis describes the development of child in preschool age and describes the speech development and speech impairments. Thesis is focused on the mediated learning experience, its principles, goals and also on the theory of structural cognitive modifiability. In the last part of thesis is describede the program of Feuerstein instrumental enrichment, which is the main part of the research. The goal of thesis is to analysed the use of the Feuerstein instrumental enrichment program in the focus on development of preschool children's communication skills.

Atmospheric haze, which builds up over South Africa including our study areas, Cape Town and the Mpumalanga Highveld under calm weather conditions, causes public concern. The scope of this study was to determine the concentration and composition of atmospheric aerosol at Khayelitsha (an urban site in the Western Cape) and Ferrobank (an industrial site in Witbank, Mpumalanga). Particulate matter was collected in Khayelitsha from 18 May 2007 to 20 July 2007 (i.e. 20 samples) using a Partisol-plus sampler and a Tapered Element Oscillating Microbalance (TEOM) sampler. Sampling took place at Ferrobank from 07 February 2008 to 11 March 2008 (6 samples) using a Partisol-plus sampler and an E-sampler. The gravimetric mass of each exposed sample was determined from pre- and post-sampling weighing. The elemental composition of the particulate matter was determined for 16 elements at Khayelitsha using Proton Induced X-ray Emission (PIXE). The concentration of the elements Al, Si, S, Cl, K, Ca, Ti, Cr, Mn, Fe, Cu, Zn, As, Br, Sn, and Pb was determined by analysing the PIXE spectra obtained. In similar manner, the elemental composition of the particulate matter was determined for 15 elements at Ferrobank (Al, Si, S, Cl, K, Ca, Ti, Cr, Mn, Fe, Cu, Zn, As, Br and Pb). The average aerosol mass concentrations for different days at the Khayelitsha site were found to vary between 8.5 μg/m3 and 124.38 μg/m3. At the Khayelitsha site on three occasions during the sampling campaign the average aerosol mass concentrations exceeded the current South African air quality standard of 75 μg/m3 over 24 h. At the Ferrobank site, there are no single days that exceeded the limit of the South African air quality standard during the sampling campaign. Enrichment factors for each element of the particles sampled with an aerodynamic diameter of less than 10 μm (PM10) samples have been calculated in order to identify their possible sources. The analysis yielded five potential sources of PM10 : soil dust, sea salt, gasoline emissions, domestic wood and coal combustion. Interestingly, enrichment factor values for the Khayelitsha samples show that sea salt constitutes a major source of emissions, while Ferrobank samples, the source apportionment by unique ratios (SPUR) indicate soil dust and coal emission are the major sources of pollution. The source apportionment at Khayelitsha shows that sea salt and biomass burning are major source of air pollution.

No description available.

v angličtině (English abstract) Tobacco male gametophyte has a strongly dehydrated cytoplasm and represents a metabolically inactive stage. Upon cytoplasm rehydration, pollen grain becomes metabolically active and after the activation is finished, the pollen tube growth through a selected pollen aperture starts. The rehydration together with metabolic activation are accompanied by the regulation of translation and post-translational modifications (mainly phosphorylation) of the existing proteins. In this Ph.D. thesis, there were identified phosphopeptides from tobacco (Nicotiana tabacum) mature pollen, pollen activated in vitro 5 min and pollen activated in vitro 30 min. The total proteins from the above male gametophyte stages were extracted. The protein extract was trypsinized and the acquired peptide mixture was enriched by MOAC (metal oxide/hydroxide affinity chromatography) with titanium dioxide matrix. The enriched fraction was subjected to liquid chromatography coupled with tandem mass spectrometry (LC- MS/MS). Totally, there were identified 471 phosphopeptides, carrying 432 exactly localized phosphorylation sites. The acquired peptide identifications were mapped to 301 phosphoproteins that were placed into 13 functional categories, dominant of which were transcription, protein synthesis,...

Au cours de la dernière décennie, les technologies permettant la numérisation d'espaces urbains ont connu un développement rapide. Des campagnes d'acquisition de données couvrant des villes entières ont été menées en utilisant des scanners LiDAR (Light Detection And Ranging) installés sur des véhicules mobiles. Les résultats de ces campagnes d'acquisition laser, représentants les bâtiments numérisés, sont des nuages de millions de points pouvant également contenir un ensemble de photographies. On s'intéresse ici à l'amélioration du nuage de points à l'aide des données présentes dans ces photographies. Cette thèse apporte plusieurs contributions notables à cette amélioration. La position et l'orientation des images acquises sont généralement connues à l'aide de dispositifs embarqués avec le scanner LiDAR, même si ces informations de positionnement sont parfois imprécises. Pour obtenir un recalage précis d'une image sur un nuage de points, nous proposons un algorithme en deux étapes, faisant appel à l'information mutuelle normalisée et aux histogrammes de gradients orientés. Cette méthode permet d'obtenir une pose précise même lorsque les estimations initiales sont très éloignées de la position et de l'orientation réelles. Une fois ces images recalées, il est possible de les utiliser pour inférer la couleur de chaque point du nuage en prenant en compte la variabilité des points de vue. Pour cela, nous nous appuyons sur la minimisation d'une énergie prenant en compte les différentes couleurs associables à un point et les couleurs présentes dans le voisinage spatial du point. Bien entendu, les différences d'illumination lors de l'acquisition des données peuvent altérer la couleur à attribuer à un point. Notamment, cette couleur peut dépendre de la présence d'ombres portées amenées à changer avec la position du soleil. Il est donc nécessaire de détecter et de corriger ces dernières. Nous proposons une nouvelle méthode qui s'appuie sur l'analyse conjointe des variations de la réflectance mesurée par le LiDAR et de la colorimétrie des points du nuage. En détectant suffisamment d'interfaces ombre/lumière nous pouvons caractériser la luminosité de la scène et la corriger pour obtenir des scènes sans ombre portée. Le dernier problème abordé par cette thèse est celui de la densification du nuage de points. En effet la densité locale du nuage de points est variable et parfois insuffisante dans certaines zones. Nous proposons une approche applicable directement par la mise en oeuvre d'un filtre bilatéral joint permettant de densifier le nuage de points en utilisant les données des images / Recent years saw a rapid development of city digitization technologies. Acquisition campaigns covering entire cities are now performed using LiDAR (Light Detection And Ranging) scanners embedded aboard mobile vehicles. These acquisition campaigns yield point clouds, composed of millions of points, representing the buildings and the streets, and may also contain a set of images of the scene. The subject developed here is the improvement of the point cloud using the information contained in the camera images. This thesis introduces several contributions to this joint improvement. The position and orientation of acquired images are usually estimated using devices embedded with the LiDAR scanner, even if this information is inaccurate. To obtain the precise registration of an image on a point cloud, we propose a two-step algorithm which uses both Mutual Information and Histograms of Oriented Gradients. The proposed method yields an accurate camera pose, even when the initial estimations are far from the real position and orientation. Once the images have been correctly registered, it is possible to use them to color each point of the cloud while using the variability of the point of view. This is done by minimizing an energy considering the different colors associated with a point and the potential colors of its neighbors. Illumination changes can also change the color assigned to a point. Notably, this color can be affected by cast shadows. These cast shadows are changing with the sun position, it is therefore necessary to detect and correct them. We propose a new method that analyzes the joint variation of the reflectance value obtained by the LiDAR and the color of the points. By detecting enough interfaces between shadow and light, we can characterize the luminance of the scene and to remove the cast shadows. The last point developed in this thesis is the densification of a point cloud. Indeed, the local density of a point cloud varies and is sometimes insufficient in certain areas. We propose a directly applicable approach to increase the density of a point cloud using multiple images

Respostas adaptativas ao estresse podem ser acompanhadas por alterações nos comportamentos emocionais, em particular relacionados com medo e ansiedade, bem como alterações na sensibilidade dolorosa. Ainda, tem sido investigado o papel do óxido nítrico em áreas encefálicas relacionadas ao comportamento defensivo. Embora várias evidências têm demonstrado que o enriquecimento ambiental promove melhora nos processos de memória, no aprendizado e em respostas nociceptivas, a relação entre o estresse crônico e as vantagens da utilização do enriquecimento ambiental ainda são pouco investigadas. O presente estudo teve como objetivo investigar se o enriquecimento ambiental promove alteração do comportamento emocional, da sensibilidade nociceptiva, bem como na imunorreatividade à nNOS no núcleo central da amígdala, na formação hipocampal e na região dorsolateral da substância cinzenta periaquedutal, em ratos submetidos ao estresse por isolamento social ou estresse crônico variado e mantidos em ambiente enriquecido ou sem enriquecimento. Ratos machos Wistar (~70g) foram divididos aleatoriamente em dois grandes grupos experimentais: Ambiente Padrão (Padrão) ou Ambiente Enriquecido (EE), mantidos por 38 dias. Cada grupo foi subdividido dependendo do tipo de estresse crônico: Controle (sem estresse), Isolamento Social (por 38 dias) e Estresse Crônico Variado (do dia 28 ao dia 37). Ao fim do tempo experimental (dia 38) os ratos foram avaliados quanto ao comportamento emocional pelos testes de labirinto em cruz elevado (LCE) e claro/escuro (TCE) e sensibilidade nociceptiva pelo teste da placa quente (a qual foi realizada em duas etapas, sendo a primeira medida no dia 0 e outra no dia 38). A eutanásia dos ratos ocorreu no dia 39, para coleta do encéfalo para análise da imunorreatividade à óxido nítrico sintase neuronal (nNOS). Levando-se em consideração o comportamento emocional e a sensibilidade nociceptiva, os diferentes tipos de estresse crônico diminuíram a porcentagem de tempo, a frequência de entrada e a exploração da extremidade dos braços abertos e na frequência de mergulho de cabeça no teste do LCE, apesar de não alterar a sensibilidade nociceptiva. Por outro lado, o enriquecimento ambiental aumentou a porcentagem de tempo, a frequência de entrada e a exploração da extremidade dos braços abertos no teste do LCE, apesar de não alterar a sensibilidade nociceptiva. Foi observado aumento da imunorreatividade à nNOS na formação hipocampal em diferentes tipos de estresse crônico. Em particular, na região de CA3 houve interação significante entre os fatores estresse por isolamento social e ambiente de manutenção. Deste modo, os resultados obtidos neste trabalho sugerem que a formação hipocampal desempenha importante função no efeito ansiogênico exercido pelos diferentes tipos de estressores crônicos (aqui representados pelo isolamento social e pelo estresse crônico variado) provavelmente pela ativação do sistema nitrérgico e sugere-se que o enriquecimento possa prevenir o comportamento do tipo ansioso. / Adaptive responses to stress may be accompanied by changes in emotional behaviors, in particular related to fear and anxiety, as well as changes in pain sensitivity. Furthermore, the role of nitric oxide in brain areas related to defensive behavior has been investigated. Although several evidences have shown that environmental enrichment improves memory processes, learning and nociceptive responses, the relationship between chronic stress and the advantages of using environmental enrichment is still poorly investigated. The present study aimed to investigate whether environmental enrichment promotes alteration of the emotional behavior, nociceptive sensitivity, as well as immunoreactivity to neuronal nitric oxide synthase (nNOS) in the central nucleus of the amygdala, hippocampal formation and dorsolateral periaqueductal gray matter in rats submitted to social isolation stress or chronic unpredictable stress and reared in enriched environment or standard environment. Male Wistar rats (~ 70g) were randomly divided into two major experimental groups: Standard Environment (Standard) or Enriched Environment (EE), maintained for 38 days. Each group was subdivided according to the type of chronic stress: Control (without stress), Social Isolation (for 38 days) and Chronic Unpredictable Stress (from day 28 to day 37). At the end of the experimental time (day 38), the rats were evaluated for emotional behavior by elevated plus maze (EPM) and light/dark box (LDBT) tests and nociceptive sensitivity by the hot plate test (which was performed in two steps , the first being measured on day 0 and the other on day 38). Euthanasia of rats occurred on day 39, to collect the brain for nNOS immunoreactivity analysis. Taking into account emotional behavior and nociceptive sensitivity, the different types of chronic stress decreased the percentage of time, the frequency of entry of the open arms, end-arm exploration and the head dipping frequency in the EPM, despite of not altering the nociceptive sensitivity. On the other hand, environmental enrichment increased the percentage of time, the frequency of entry of the open arms and the end arm-exploration in the EPM test, although it did not alter the nociceptive sensitivity. Increased immunoreactivity to nNOS in hippocampal formation was observed in different types of chronic stress. In particular, in the CA3 region there was a significant interaction between stress factors due to social isolation and maintenance environment. Thus, the results obtained in this study suggest that hippocampal formation plays an important role in the anxiogenic effect exerted by the different types of chronic stressors (represented here by social isolation and by chronic chronic stress) probably due to the activation of the nitrergic system and it is suggested that environmental enrichment can prevent of anxiety-like behavior.

A Bioacoustica oferece ferramentas úteis para diversas ciências e práticas na utilização acadêmica, científica, aplicada e conservação da vida animal. Nossa hipótese é que as chamadas naturais, como o rugido de longo alcance do bugios preto, atuam como elemento de enriquecimento ambiental, se a sua execução aumenta a exibição de um comportamento natural em cativeiro. O bugio faz um uso diário do rugido e sua emissão é conhecida por evocar respostas de outros bandos de bugio em vida livre. Para testar a eficácia dos rugidos em melhorar o bem-estar foi realizado um experimento no Bosque e Zoológico Fábio Barreto, no município de Ribeirão Preto, onde treze indivíduos (6 fêmeas e 7 machos) viviam em cinco recintos adjacentes. Eles tiveram contato visual e acústico entre eles e com outras duas espécies de primatas e outros grupos de vida livre de bugio preto. Adotoamos o delineamento experimental ABABA, em que fases controle A foram intercaladas com o tratamento (playback de rugido) nas fases B, duas vezes ao dia (manhã e tarde). Na primeira fase B, o estímulo foi sempre tocado no mesmo horário (previsível) e, na segunda fase tratamento, horários aleatórios foram adotados (imprevisível). Nós avaliamos as alterações na duração relativa de 6 comportamentos entre as fases A e B, entre sexo e em ambos os períodos, com o teste ANOVA de medidas repetidas. Nós notamos um aumento no comportamento natural da espécie em fases de tratamento, como alerta, atenção dirigida, e interações sociais, juntamente com a diminuição de comportamentos estereotipados, sugerindo melhoria do bem-estar. As emissões naturais de rugidos também aumentaram. Concluímos que ambos os tratamentos, previsível e imprevisível, melhoraram o bem-estar e propomos a utilização da reprodução de chamadas específicas de especies como uma nova categoria de enriquecimento ambiental. A resposta dos outros primatas nos levou a concluir que houve um efeito sobre animais vizinhos que pode ser benéfico ou prejudicial de acordo com a espécie e a condição psicológica dos animais. / The Bioacoustics offers useful tools for diverse academic, scientific and applied sciences and practices in use and conservation of animal life. We hypothesized that natural call, as the loud roar of the black-howler monkey, would act as environmental enrichment element if its execution increased the display of natural behavior in captivity. The howler-monkey makes an everyday use of the roar and its emission is known to evoke free-living howler-monkeys to call back in response. To test the effectiveness of roars in improving welfare we conducted an experiment in Bosque e Zoológico Fábio Barreto, in the municipality of Ribeirão Preto where thirteen individuals (6 females and 7 males) were living in five adjacent enclosures. They had visual and acoustic contact between them and with two other primate species and a free-living black-howler monkey group. We adopted the ABABA experimental design, in which control phases A were interspersed with treatment (roar playback) phases B, two times a day (morning and afternoon). In the first B phase, the stimulus was always played at the same time (predictable) and, in the other different random schedules were adopted (unpredictable). We evaluate changes in relative duration of 6 behaviors from A to B phases, between sex and in both periods with a repetitive factorial ANOVA. We reported increase in natural behavior of the species in treatment phases, as alert, oriented attention, and social interactions, coupled with the decrease of stereotyped behaviors, suggesting welfare improvement. Thus, natural emissions of roars also increased. We concluded that both predictable and unpredictable routines improved welfare and we propose the use of playback of specie-specific calls as a new category of environmental enrichment. The answer of the other primates led us to conclude that there was a neighborhood effect that can be beneficial or harmful according to the species and the psychological condition of the animals.