Development of 3D super-resolution tomographic STED microscopy and its application to studies on bone resorption

In this doctoral thesis, a tomographic STED microscopy technique for 3D super-resolution imaging was developed and utilized to observebone remodeling processes. To improve upon existing methods, wehave used a tomographic approach using a commercially available stimulated emission depletion (STED) microscope. A certain region of interest (ROI) was observed at two oblique angles: one at a standard inverted configuration from below (bottom view) and another from the side (side view) via a micro-mirror positioned close to the ROI. The two viewing angles were reconstructed into a final tomogram. The technique, named as tomographic STED microscopy, was able to achieve an axial resolution of approximately 70 nm on microtubule structures in a fixed biological specimen. High resolution imaging of osteoclasts (OCs) that are actively resorbing bone was achieved by creating an optically transparent coating on a microscope coverglass that imitates a fractured bone surface. 2D super-resolution STED microscopy on the bone layer showed approximately 60 nm of lateral resolution on a resorption associated organelle allowing these structures to be imaged with super-resolution microscopy for the first time. The developed tomographic STED microscopy technique was further applied to study resorption mechanisms of OCs cultured on the bone coating. The technique revealed actin cytoskeleton with specific structures, comet-tails, some of which were facing upwards and some others were facing downwards. This, in our opinion, indicated that during bone resorption, an involvement of the actin cytoskeleton in vesicular exocytosis and endocytosis is present. The application of tomographic STED microscopy in bone biology demonstrated that 3D super-resolution techniques can provide new insights into biological 3D nano-structures that are beyond the diffraction-limit when the optical constraints of super-resolution imaging are carefully taken into account.

Atividade amilol��tica e qualidade fisiol��gica de sementes armazenadas de milho super doce tratadas com ��cido giber��lico

O uso de reguladores de crescimento na fase de germina����o melhora o desempenho das pl��ntulas, acelerando a velocidade de emerg��ncia e real��ando o potencial das sementes de v��rias esp��cies, mesmo sob condi����es adversas. Este trabalho teve como objetivo avaliar a influ��ncia do ��cido giber��lico na atividade amilol��tica e no vigor de sementes armazenadas de milho super doce. O experimento foi conduzido nos Laborat��rios de An��lise de Sementes do Departamento de Produ����o Vegetal/FCA e no Laborat��rio de Bioqu��mica de Plantas do Departamento de Qu��mica e Bioqu��mica/IB da Universidade Estadual Paulista (UNESP/Botucatu), entre os meses de julho e setembro de 2001, onde foram feitas as avalia����es da qualidade fisiol��gica, atrav��s dos testes de germina����o, vigor e bioqu��micos. Sementes de milho super doce da cultivar DO-04, foram acondicionadas em sacos de papel e armazenadas por oito meses em c��mara seca (40% UR). Ap��s este per��odo, foram colocadas para germinar em rolos de papel toalha, embebidos com GA3 nas concentra����es zero; 50; 100; 150 e 200mg.L-1. Foram avaliadas a germina����o, vigor e atividade amilol��tica das sementes. As sementes submetidas �� pr��-embebi����o em solu����o de 50mg.L-1 de ��cido giber��lico, apresentaram maior germina����o e vigor, menor teor de prote��nas totais e maior atividade amilol��tica.

Matura����o de sementes de milho-doce: grupo Super Doce

O experimento teve como objetivo estudar o processo de matura����o de sementes de milho doce da cultivar BR 400 (Super Doce). A primeira colheita das sementes foi realizada 27 dias ap��s a flora����o (DAF), a qual foi considerada quando aproximadamente 50% das plantas apresentavam florescimento feminino. As demais colheitas foram realizadas em intervalos de sete dias, at�� 90 DAF. Imediatamente ap��s a colheita, procedeu-se �� debulha manual das sementes, determinando-se o grau de umidade e o peso da mat��ria seca; avaliou se tamb��m, visualmente, a presen��a da camada negra. Ap��s a ��ltima colheita, foi determinada a qualidade fisiol��gica das sementes por meio dos testes de germina����o, frio modificado, envelhecimento acelerado e condutividade el��trica. As sementes de milho-doce atingiram o m��ximo de mat��ria seca (maturidade de massa) aproximadamente aos 41 DAF, sendo que o m��ximo de germina����o e vigor (maturidade fisiol��gica) ocorreu no per��odo compreendido entre 48 e 76 DAF. Todas as sementes apresentaram camada negra aos 55 DAF, sendo esta uma caracter��stica visual eficiente para a identifica����o da maturidade fisiol��gica das sementes.

Tratamento fungicida em semente de milho super-doce

Neste trabalho, procurou-se avaliar a influ��ncia do tratamento fungicida no potencial fisiol��gico e na sanidade de sementes de milho h��brido super-doce DO-04, separadas por diferen��as quanto ao formato. Foi utilizado delineamento inteiramente casualizado, em esquema fatorial 2 x 3 x 4, compreendendo dois formatos de semente (retidas em peneira 15 x ��", representando as sementes arredondadas, e em peneira 20, representando as achatadas), tr��s lotes e quatro tratamentos fungicidas (testemunha, Fludioxonil, Captan TS + Thiabendazol e Captan TS + Thiram PS). Avaliaram-se a germina����o, o vigor (velocidade de germina����o, teste de frio e envelhecimento acelerado) e a sanidade de semente. Foi verificado que as sementes arredondadas de milho h��brido super-doce DO-04 apresentam germina����o e vigor mais elevados, em compara����o ��s achatadas; os tratamentos fungicidas favorecem o vigor, sendo as combina����es Captan + Thiabendazol e Captan + Thiram eficientes para o controle de F. moniliforme, Cephalosporium sp. e Penicillium sp..

The debt crisis: a re-appraisal

The 1980s' debt crisis is a landmark in developing economies' growth and stabilization. According to the most quoted empirical articles, external shocks and vicissitudes gave rise to crisis just because of delays in stabilization policies, engendered by internal conflicts and institutional immaturity. I review some of these papers, and find out some problems - in the measurement of shocks and foreign indebtedness, namely - whose corrections lead to opposite results: external shocks and foreign indebtedness explain that crisis regardless of domestic policies. At the same time, the strong correlation of income distribution to terms of trade changes and foreign indebtedness suggest that inequality may have contributed differently to that crisis: either through an economic channel, or through a political channel based on delays in reforms.

Entry mode choice of Finnish startups entering Chinese market in tablet industry

Since the implement of opening policy, the overall economy of China has maintained rapid and stable development, which has now makes China become the world's second largest economy. China, it is to become the largest overseas market for many large global enterprises from various industries, this naturally also includes the Tablet PC industry that raised in recent years. The purpose of this thesis is to analyze different internal and external factors that influence the entry mode choices of Finnish SMEs in tablet industry entering Chinese market. The goal is to find out the suitable entry modes for the Finnish tablet or other relevant SMEs entering Chinese market. Qualitative analysis is the main research method in empirical part of this study. The interviews were carried out with the case company and other two Finnish business organizations in China. The result of the study indicated that the internal resource and external business environment affect the entry modes choices much more than other factors for SMES. The exporting mode and sales subsidiary could be a better choice for SMEs entering Chinese market. Furthermore, firms should fully learn the Chinese market combine with its own background before making decisions.

Business process re-engineering -menetelm��t myyntiprosessin tehokkuuden parantamisessa (Case-yritys)

T��m��n Pro Gradu-tutkielman tavoite on tutkia Business Process Re-engineering menetelmi�� myyntiprosessien tehostamisessa. Tutkimuksen teoreettinen viiteke-hys rakentuu myynninjohtamisen, myyntiprosessien ja Business Process Mana-gementin ja Business Process Re-enineeringin ymp��rille. IT-j��rjestelm��t ovat my��s oleellinen osa-alue tutkimuksen kannalta ja niiden osuutta kuvataan niin myyntiprosesseissa kuin Business Process Re-engineering -menetelmien yhtey-dess��. Tutkielmassa perehdyt����n aikaisempaan tutkimusmateriaaliin ja akateemiseen kirjallisuuteen yll��mainituilla osa-alueilla. Tavoitteena on l��yt���� aikaisempia tutki-muksia myyntiprosessien tehostamisesta ja BPR:n roolista n��iss�� tapauksissa. My��s myynninjohtamisen vaikutusta tehokkaaseen myyntiprosessiin tutkitaan, kuten my��s IT-j��rjestelmien erilaisia rooleja tehokkaissa myyntiprosesseissa. Tutkielman empiirinen osio on kvalitatiivinen Case-tutkimus er����ss�� rahoitusalan yrityksess��. Tutkimus tehd����n haastattelemalla myyntihenkil��st���� ja esimiehi��. Lis��ksi analysoidaan yrityksen myyntiprosessiin liittyv���� muuta materiaalia. Case-tutkimuksen tuloksia peilataan aiempaan akateemiseen tutkimukseen ja tuloksista pyrit����n l��yt��m����n ratkaisuja, miten BPR -menetelmill�� voidaan tehostaa yrityksen myyntiprosessia.

Did ISI fail and is neoliberalism the answer for Latin America? Re-assessing common wisdom regarding economic policies in the region

The second phase of Import Substituting Industrialization, commonly known as ISI2, involved the move in Latin America to "heavy" industrialization, from around 1950-80. This period of economic history has been reviled on both the Left and the Right as being one of either heightened dependency or one demonstrating the clear failure of state intervention in the economy. In this research note, a basic statistical analysis is used to back up other descriptive claims that the ISI2 period was rather one of mixed success, with macroeconomic volatility accompanying great progress in GDP and manufacturing growth. In a sense, the ISI2 period succeeded in industrializing the large economies of the period, and contrasts favorably with the record of the succeeding paradigm of neoliberalism. This research note seeks to raise questions about the way we look at the historical period of ISI2, and suggests that a more open-minded perspective could lead to a more effective and sustainable political economy paradigm for the region in the future.

Bioimage informatics in STED super-resolution microscopy

Optical microscopy is living its renaissance. The diffraction limit, although still physically true, plays a minor role in the achievable resolution in far-field fluorescence microscopy. Super-resolution techniques enable fluorescence microscopy at nearly molecular resolution. Modern (super-resolution) microscopy methods rely strongly on software. Software tools are needed all the way from data acquisition, data storage, image reconstruction, restoration and alignment, to quantitative image analysis and image visualization. These tools play a key role in all aspects of microscopy today ��� and their importance in the coming years is certainly going to increase, when microscopy little-by-little transitions from single cells into more complex and even living model systems. In this thesis, a series of bioimage informatics software tools are introduced for STED super-resolution microscopy. Tomographic reconstruction software, coupled with a novel image acquisition method STED< is shown to enable axial (3D) super-resolution imaging in a standard 2D-STED microscope. Software tools are introduced for STED super-resolution correlative imaging with transmission electron microscopes or atomic force microscopes. A novel method for automatically ranking image quality within microscope image datasets is introduced, and it is utilized to for example select the best images in a STED microscope image dataset.

(Re)descrevendo Foucault: com Rorty, contra Rorty

O artigo prop��e uma interlocu����o entre o fil��sofo franc��s Michel Foucault e o fil��sofo norte-americano Richard Rorty. Apresenta a descri����o que Rorty realizou do colega franc��s. Analisa essa leitura e oferece, a partir do pr��prio Foucault, uma interpreta����o alternativa, que aponta para algumas imprecis��es cometidas por Rorty, em sua interpreta����o. Conclui com um coment��rio sobre a conversa����o proposta.