A resistência bacteriana a antibióticos é um grave e crescente problema de saúde pública de âmbito mundial. O principal, e mais eficiente, mecanismo de resistência aos &#946-lactâmicos em bacilos Gram-negativos é a produção de &#946-lactamases, que possuem a capacidade de hidrolisar o anel &#946-lactâmicos e consequentemente inativar essa classe de antibióticos. Vale ressaltar, que atualmente os antibióticos &#946-lactâmicos são os mais utilizados clinicamente, particularmente em infecções graves. Dentre as &#946-lactamases existentes destacam-se as carbapenemases, enzimas capazes de inativar a maioria dos antibióticos &#946-lactâmicos. Uma grande preocupação é o fato dessas enzimas, em sua maioria, serem codificadas por plasmídeos, o que propicia a disseminação desses genes de resistência; portanto, é de extrema importância a realização de um rápido e efetivo monitoramento da presença de patógenos portadores desses genes de resistência, para que assim se possa prevenir a disseminação desses determinantes. Foram incluídos neste estudo 230 amostras únicas de Acinetobacter e Pseudomonas aeruginosa resistentes a imipenem detectados em pacientes internados em hospitais privados da cidade de São Paulo durante o período de fevereiro a outubro de 2013. As amostras foram avaliadas quanto à hidrólise de imipenem por espectrofotometria, quanto à presença de genes de carbapenemases por PCR e sequenciamento, e quanto à clonalidade por eletroforese em campos pulsados (PFGE) ou ERIC-PCR. Foram realizados ensaios de conjugação, transformação e sequenciamento completo de plasmídeos. Dentre as amostras de Acinetobacter spp. 80% (88) foram capazes de hidrolisar o imipenem. Dentre esses 76,1% (67) foram positivos para blaOXA-51-like, 19,3% (17) foram positivos para blaOXA-72. blaOXA-23, blaOXA-482 e blaIMP-1 foram detectados isoladamente em isolados distintos. O gene blaIMP-1 foi detectado em A. ursingii inserido em integron de classe 1 e representa a primeira descrição no Brasil. Uma nova carbapenemase OXA-482-like foi detectada em A. baumanii. Utilizando-se ERIC-PCR, observou-se uma grande diversidade de grupos clonais, com o máximo de quatro isolados por grupo. Dentre as amostras de P. aeruginosa, apenas 35,3% foram capazes de hidrolisar o imipenem. Dessas amostras, 14 possuíam o gene blaSPM-1, e isolados únicos possuíam, individualmente, os genes blaIMP, blaVIM, blaKPC-2 ou blaGES-23. O gene blaKPC-2 foi detectado inserido em contexto genético diferente dos descritos anteriormente, em plasmídeo IncU de 32 Kb, mobilizável, mas não conjugativo. Esta é a primeira descrição da sequencia completa de plasmídeo albergando o gene blaKPC-2 em P. aeruginosa no Brasil. Nas demais amostras (20) com atividade hidrolítica, não foram detectados genes de carbapenemase conhecidos, o que sugere a presença de genes de carbapenemase ainda não descritos. Em três amostras foi possível obter transformantes com plasmídeos, resistentes a carbapenêmicos. As amostras com blaSPM-1 apresentaram perfis de PFGE estreitamente relacionados. Em contraste, os perfis de PFGE das amostras com potenciais novas carbapenemases apresentaram índice de similaridade de Dice inferior ix a 80%, evidenciando grande diversidade clonal. Nossos achados evidenciam que a carbapenemase não intrínseca predominante em Acinetobacterem hospitais privados da cidade de São Paulo é OXA-72, e em hospitais privados há uma grande diversidade clonal. Em P. aeruginosa, a carbapenemase predominante é SPM-1, cuja disseminação é mediada por um único clone. Há potencialmente um número significativo de novas carbapenemases em Acinetobacter e P. aeruginosa, algumas delas mediadas por plasmídeos. / Bacterial resistance to antibiotics is a serious and growing public health problem worldwide. The main and most efficient mechanism of resistance to &#946-lactams in Gram-negative bacilli is the production of &#946-lactamases, which have the ability to hydrolyze the &#946-lactam ring and consequently inactivate this class of antibiotics. It is worth mentioning that currently &#946-lactam antibiotics are the most used clinically, particularly in severe infections. Among the existing &#946-lactamases, carbapenemases are capable of inactivating most &#946-lactam antibiotics. A major concern is that these enzymes are mostly encoded by plasmids, which facilitates the spread of these resistance genes; therefore, it is of extreme importance to carry out a rapid and effective monitoring of the presence of pathogens bearing these resistance genes, in order to prevent the dissemination of these determinants. This study included 230 unique samples of imipenem-resistant Acinetobacterand Pseudomonas aeruginosa detected in patients hospitalized in private hospitals in the city of São Paulo during the period from February to October 2013. The samples were evaluated for the imipenem hydrolysis by spectrophotometry, the presence of carbapenemase genes by PCR and sequencing, and concerning clonality by pulsed field electrophoresis (PFGE) or ERIC-PCR. Conjugation, transformation and complete sequencing of plasmids were performed. Among Acinetobacter spp. samples, 80% (88) were able to hydrolyze imipenem. Among these, 76.1% (67) were positive for blaOXA-51-like genes and 19.3% (17) were positive for blaOXA-72. The blaOXA-23, blaOXA-482 and blaIMP-1 genes were detected alone in distinct isolates. The blaIMP-1 gene was detected in A. ursingii inserted in class 1 integron and represents the first description in Brazil. A novel OXA-482-like carbapenemase was detected in A. baumanii. Using ERIC-PCR, a great diversity of clonal groups was observed, with a maximum of four isolates per group. Among P. aeruginosa samples, only 35.3% were able to hydrolyze imipenem. Of these samples, 14 had the blaSPM-1 gene, and single isolates individually possessed the blaIMP, blaVIM, blaKPC-2 or blaGES-23 genes. The blaKPC-2 gene was found inserted in a genetic context different from those described previously, in a mobilizable, but not conjugative, 32 Kb IncU plasmid. This is the first description of the complete nucleotide sequence of a plasmid harboring the blaKPC-2 gene in P. aeruginosa in Brazil. In the remaining samples (20) with hydrolytic activity, no known carbapenemase genes were detected, suggesting the presence of carbapenemase genes not yet described. In three samples it was possible to obtain transformants with plasmids, resistant to carbapenems. Samples with blaSPM-1 showed closely related PFGE profiles. In contrast, the PFGE profiles of the samples with potential new carbapenemases showed Dice similarity index lower than 80%, evidencing a great clonal diversity. Our findings show that the predominant non-intrinsic carbapenemase in Acinetobacter in the city of São Paulo is OXA-72, and in private hospitals there is great clonal diversity. In P. aeruginosa, the predominant carbapenemase is SPM-1, the spread of this enzyme is mediated by a single clone. There are potentially a significant number of new carbapenemases in Acinetobacter and P. aeruginosa, some of them plasmid mediated.

A resistência bacteriana a antibióticos é um grave e crescente problema de saúde pública de âmbito mundial. O principal, e mais eficiente, mecanismo de resistência aos &#946-lactâmicos em bacilos Gram-negativos é a produção de &#946-lactamases, que possuem a capacidade de hidrolisar o anel &#946-lactâmicos e consequentemente inativar essa classe de antibióticos. Vale ressaltar, que atualmente os antibióticos &#946-lactâmicos são os mais utilizados clinicamente, particularmente em infecções graves. Dentre as &#946-lactamases existentes destacam-se as carbapenemases, enzimas capazes de inativar a maioria dos antibióticos &#946-lactâmicos. Uma grande preocupação é o fato dessas enzimas, em sua maioria, serem codificadas por plasmídeos, o que propicia a disseminação desses genes de resistência; portanto, é de extrema importância a realização de um rápido e efetivo monitoramento da presença de patógenos portadores desses genes de resistência, para que assim se possa prevenir a disseminação desses determinantes. Foram incluídos neste estudo 230 amostras únicas de Acinetobacter e Pseudomonas aeruginosa resistentes a imipenem detectados em pacientes internados em hospitais privados da cidade de São Paulo durante o período de fevereiro a outubro de 2013. As amostras foram avaliadas quanto à hidrólise de imipenem por espectrofotometria, quanto à presença de genes de carbapenemases por PCR e sequenciamento, e quanto à clonalidade por eletroforese em campos pulsados (PFGE) ou ERIC-PCR. Foram realizados ensaios de conjugação, transformação e sequenciamento completo de plasmídeos. Dentre as amostras de Acinetobacter spp. 80% (88) foram capazes de hidrolisar o imipenem. Dentre esses 76,1% (67) foram positivos para blaOXA-51-like, 19,3% (17) foram positivos para blaOXA-72. blaOXA-23, blaOXA-482 e blaIMP-1 foram detectados isoladamente em isolados distintos. O gene blaIMP-1 foi detectado em A. ursingii inserido em integron de classe 1 e representa a primeira descrição no Brasil. Uma nova carbapenemase OXA-482-like foi detectada em A. baumanii. Utilizando-se ERIC-PCR, observou-se uma grande diversidade de grupos clonais, com o máximo de quatro isolados por grupo. Dentre as amostras de P. aeruginosa, apenas 35,3% foram capazes de hidrolisar o imipenem. Dessas amostras, 14 possuíam o gene blaSPM-1, e isolados únicos possuíam, individualmente, os genes blaIMP, blaVIM, blaKPC-2 ou blaGES-23. O gene blaKPC-2 foi detectado inserido em contexto genético diferente dos descritos anteriormente, em plasmídeo IncU de 32 Kb, mobilizável, mas não conjugativo. Esta é a primeira descrição da sequencia completa de plasmídeo albergando o gene blaKPC-2 em P. aeruginosa no Brasil. Nas demais amostras (20) com atividade hidrolítica, não foram detectados genes de carbapenemase conhecidos, o que sugere a presença de genes de carbapenemase ainda não descritos. Em três amostras foi possível obter transformantes com plasmídeos, resistentes a carbapenêmicos. As amostras com blaSPM-1 apresentaram perfis de PFGE estreitamente relacionados. Em contraste, os perfis de PFGE das amostras com potenciais novas carbapenemases apresentaram índice de similaridade de Dice inferior ix a 80%, evidenciando grande diversidade clonal. Nossos achados evidenciam que a carbapenemase não intrínseca predominante em Acinetobacterem hospitais privados da cidade de São Paulo é OXA-72, e em hospitais privados há uma grande diversidade clonal. Em P. aeruginosa, a carbapenemase predominante é SPM-1, cuja disseminação é mediada por um único clone. Há potencialmente um número significativo de novas carbapenemases em Acinetobacter e P. aeruginosa, algumas delas mediadas por plasmídeos. / Bacterial resistance to antibiotics is a serious and growing public health problem worldwide. The main and most efficient mechanism of resistance to &#946-lactams in Gram-negative bacilli is the production of &#946-lactamases, which have the ability to hydrolyze the &#946-lactam ring and consequently inactivate this class of antibiotics. It is worth mentioning that currently &#946-lactam antibiotics are the most used clinically, particularly in severe infections. Among the existing &#946-lactamases, carbapenemases are capable of inactivating most &#946-lactam antibiotics. A major concern is that these enzymes are mostly encoded by plasmids, which facilitates the spread of these resistance genes; therefore, it is of extreme importance to carry out a rapid and effective monitoring of the presence of pathogens bearing these resistance genes, in order to prevent the dissemination of these determinants. This study included 230 unique samples of imipenem-resistant Acinetobacterand Pseudomonas aeruginosa detected in patients hospitalized in private hospitals in the city of São Paulo during the period from February to October 2013. The samples were evaluated for the imipenem hydrolysis by spectrophotometry, the presence of carbapenemase genes by PCR and sequencing, and concerning clonality by pulsed field electrophoresis (PFGE) or ERIC-PCR. Conjugation, transformation and complete sequencing of plasmids were performed. Among Acinetobacter spp. samples, 80% (88) were able to hydrolyze imipenem. Among these, 76.1% (67) were positive for blaOXA-51-like genes and 19.3% (17) were positive for blaOXA-72. The blaOXA-23, blaOXA-482 and blaIMP-1 genes were detected alone in distinct isolates. The blaIMP-1 gene was detected in A. ursingii inserted in class 1 integron and represents the first description in Brazil. A novel OXA-482-like carbapenemase was detected in A. baumanii. Using ERIC-PCR, a great diversity of clonal groups was observed, with a maximum of four isolates per group. Among P. aeruginosa samples, only 35.3% were able to hydrolyze imipenem. Of these samples, 14 had the blaSPM-1 gene, and single isolates individually possessed the blaIMP, blaVIM, blaKPC-2 or blaGES-23 genes. The blaKPC-2 gene was found inserted in a genetic context different from those described previously, in a mobilizable, but not conjugative, 32 Kb IncU plasmid. This is the first description of the complete nucleotide sequence of a plasmid harboring the blaKPC-2 gene in P. aeruginosa in Brazil. In the remaining samples (20) with hydrolytic activity, no known carbapenemase genes were detected, suggesting the presence of carbapenemase genes not yet described. In three samples it was possible to obtain transformants with plasmids, resistant to carbapenems. Samples with blaSPM-1 showed closely related PFGE profiles. In contrast, the PFGE profiles of the samples with potential new carbapenemases showed Dice similarity index lower than 80%, evidencing a great clonal diversity. Our findings show that the predominant non-intrinsic carbapenemase in Acinetobacter in the city of São Paulo is OXA-72, and in private hospitals there is great clonal diversity. In P. aeruginosa, the predominant carbapenemase is SPM-1, the spread of this enzyme is mediated by a single clone. There are potentially a significant number of new carbapenemases in Acinetobacter and P. aeruginosa, some of them plasmid mediated.

The thesis is focused on development of scanning probe microscope systems, especially development and implementation of quartz resonator probes. The quartz resonator probes, compared to the standard silicon cantilevers, have several advantages. It is in particular their mechanical properties and possibility of direct electrical readout of the deflection signal. Due to the fact, the probes are easy to implement even into more complex SPM systems. The thesis deals with development of universal and open SPM control system electronics. The electronics consist of the commercial SPM control and oscillation units, the development of the other electronic parts (such as the high voltage amplifier and the preamplifier units) is described in the thesis. Further, the thesis reports on development of the qPlus UHV LT SPM microscope system that was carried out at Universität Hamburg. Part of it was development of the qPlus preamplifier able to operate at liquid helium temperature. The third topic of the thesis is the implementation of qPlus technology into the UHV VT SPM microscope suitable to operate in situ with a scanning electron microscope. The qPlus sensors and the universal UHV preamplifier were designed and manufactured. Test measurements were conducted on all of the developed systems.

<p>Functional Magnetic Resonance Imaging (fMRI) is a neuroimaging technique used to study brain functionality to enhance our understanding of the brain. This technique is based on MRI, a painless, noninvasive image acquisition method without harmful radiation. Small local blood oxygenation changes which are reflected as small intensity changes in the MR images are utilized to locate the active brain areas. Radio frequency pulses and a strong static magnetic field are used to measure the correlation between the physical changes in the brain and the mental functioning during the performance of cognitive tasks.</p><p>This master thesis presents approaches for the analysis of fMRI data. The constrained Canonical Correlation Analysis (CCA) which is able to exploit the spatio-temporal nature of an active area is presented and tested on real human fMRI data. The actual distribution of active brain voxels is not known in the case of real human data. To evaluate the performance of the diagnostic algorithms applied to real human data, a modified Receiver Operating Characteristics (modified ROC) which deals with this lack of knowledge is presented. The tests on real human data reveal the better detection efficiency with the constrained CCA algorithm.</p><p>A second aim of this thesis was to implement the promising technique of constrained CCA into the software environment SPM. To implement the constrained CCA algorithms into the fMRI part of SPM2, a toolbox containing Matlab functions has been programmed for the further use by neurological scientists. The new SPM functionalities to exploit the spatial extent of the active regions with CCA are presented and tested.</p>

Image registration is the process of aligning two images such that their mutual features overlap. This is of great importance in several medical applications. In 2008 a novel method for simultaneous T1, T2 and proton density quantification was suggested. The method is in the field of quantitative Magnetic Resonance Imaging or qMRI. In qMRI parameters are quantified by a pixel-to-pixel fit of the image intensity as a function of different MR scanner settings. The quantification depends on several volumes of different intensities to be aligned. If a patient moves during the data aquisition the datasets will not be aligned and the results are degraded due to this. Since the quantification takes several minutes there is a considerable risk of patient movements. In this master thesis three image registration methods are presented and a comparison in robustness and speed was made. The phase based algorithm was suited for this problem and limited to finding rigid motion. The other two registration algorithms, originating from the Statistical Parametrical Mapping, SPM, package, were used as references. The result shows that the pixel-to-pixel fit is greatly improved in the datasets with found motion. In the comparison between the different methods the phase based algorithm turned out to be both the fastest and the most robust method.

Functional Magnetic Resonance Imaging (fMRI) is a neuroimaging technique used to study brain functionality to enhance our understanding of the brain. This technique is based on MRI, a painless, noninvasive image acquisition method without harmful radiation. Small local blood oxygenation changes which are reflected as small intensity changes in the MR images are utilized to locate the active brain areas. Radio frequency pulses and a strong static magnetic field are used to measure the correlation between the physical changes in the brain and the mental functioning during the performance of cognitive tasks. This master thesis presents approaches for the analysis of fMRI data. The constrained Canonical Correlation Analysis (CCA) which is able to exploit the spatio-temporal nature of an active area is presented and tested on real human fMRI data. The actual distribution of active brain voxels is not known in the case of real human data. To evaluate the performance of the diagnostic algorithms applied to real human data, a modified Receiver Operating Characteristics (modified ROC) which deals with this lack of knowledge is presented. The tests on real human data reveal the better detection efficiency with the constrained CCA algorithm. A second aim of this thesis was to implement the promising technique of constrained CCA into the software environment SPM. To implement the constrained CCA algorithms into the fMRI part of SPM2, a toolbox containing Matlab functions has been programmed for the further use by neurological scientists. The new SPM functionalities to exploit the spatial extent of the active regions with CCA are presented and tested.

Valstybinės švietimo strategijos 2003–2012 metų nuostatose, patvirtintose Lietuvos Respublikos Seimo 2003 m. liepos 4 d. nutarimu Nr. IX-1700 (Žin., 2003, Nr.71-3216), nurodoma, kad siekiant užtikrinti švietimo plėtotės prieinamumą, tęstinumą ir socialinį teisingumą, turi būti užtikrinamos socialiai teisingos mokymosi ir studijų sąlygos, sukuriamos galimybės veiksmingai ugdyti įvairių gebėjimų ir poreikių vaikus. Atlikus mokslinės literatūros analizę, galima pastebėti, kad apie specialiųjų poreikių asmenų integraciją visuomenėje yra rašę nemažai autorių: A. Galkienė, V. Rajeckas, G. Varoneckas, D. Žemaitytė, M. Barkauskaitė, J. Ambrukaitis, V. Gudonis ir kiti. Tačiau nepavyko aptikti duomenų apie integruoto ugdymo strateginį aspektą. Todėl moksliniame darbe norėta tai išsiaiškinti. Šiame kontekste formuluojama magistrinio darbo tyrimo hipotezė: Jei galima identifikuoti specialiųjų poreikių vaikų integruoto ugdymo tendencijas ir problemas, tai galima parengti jų jų ugdymo bendrojo lavinimo mokyklose strategines gaires. Tyrimo objektas: specialiųjų poreikių vaikų integruotas ugdymas bendrojo lavinimo mokyklose strateginiu aspektu. Tyrimo tikslas: sumodeliuoti specialiųjų poreikių vaikų ugdymo bendrojo lavinimo mokyklose strategines gaires. Tyrimo uždaviniai: aptarti specialiųjų poreikių vaikų ugdymo Lietuvoje tendencijas; išryškinti specialiųjų poreikių vaikų integruoto ugdymo bendrojo lavinimo mokyklose problemas; empiriškai ištirti specialiųjų... [toliau žr. visą tekstą] / In the 2003–2012 years regulations of national educational strategy which are confirmed by Seimas of the republic of Lithuania decision Number IX-1700 (Žin., 2003, Nr.71-3216) in 2003 July 4, it is stated that if you want to vouch accessibility, succession and social justice of educational development you must confirm the socially just learning conditions, create an effective opportunity to develop children who have different abilities and needs. According to analysis of nonfiction, it is possible to state that there are many authors who have written about integration of people who have special needs. They are A. Galkienė, V. Rajeckas, G. Varoneckas, D. Žemaitytė, M. Barkauskaitė, J. Ambrukaitis, V. Gudonis and the others. However, it was unsuccessful to find information about strategic aspect of integrated education. So it was tried to solve this in the work. The hypothesis of this postgraduate studies work is: if it is possible to identify integrated educational tendency and problems of children who have special needs, so it is possible to prepare strategic guideline of their development in comprehensive schools. An object of this research is integrated education of children who have special needs in comprehensive schools. The aim of research is to design the strategic guideline of children development that has special needs in comprehensive schools. The goals of this research are to discuss the tendencies of children, who have special needs, development in Lithuania; to... [to full text]

This thesis presents a battery electric bus energy consumption model (ECONS-M) coupled with an electrochemical battery capacity fade model (CFM). The underlying goals of the project were to develop analytical tools to support the integration of battery electric buses. ECONS-M projects the operating costs of electric bus and the potential emission reductions compared to diesel vehicles for a chosen transit route. CFM aims to predict the battery pack lifetime expected under the specific driving conditions of the route. A case study was run for a transit route in Victoria, BC chosen as a candidate to deploy a 2013 BYD electric bus. The novelty of this work mainly lays in its application to battery electric buses, as well as in the coupling of the ECONS-M and the electrochemical model to predict how long the batteries can last if the electric bus is deployed on a specific transit route everyday. An in-depot charging strategy is the only strategy examined in this thesis due to the charging rate limitations of the electrochemical model. The ECONS-M is currently being utilized in industry for the preparations of Phase I and II of the Pan-Canadian Electric Bus Demonstration & Integration Trial led by the Canadian Urban Transit Research and Innovation Consortium (CUTRIC). This project aims to deploy up to 20 battery electric buses for phase I and 60 electric buses for phase II across Canada to support the standardization of overhead fast chargers and in-depot chargers, which in a first in the world. At this time, the developed CFM can not support any final claims due to the lack of electrochemical data in the literature for the high capacity lithium-ion cells used in electric buses. This opens the door to more research in the ageing testing of batteries for heavy-duty applications. / Graduate

The main task of this work is to design an algorithm for suppressing unwanted noise and artifacts in fMRI data using the analysis of independent components and multi-echo data. The theoretical part deals with the basic principles of magnetic resonance, including construction and image data processing. The practical part presents a pilot design of a method inspired by a professional publication in the Matlab software environment, where this design is subsequently tested on real fMRI data provided by the Laboratory of Multimodal and Functional Imaging, CEITEC MU.

Thesis discusses the principles and methodology for measuring functional magnetic resonance imaging (fMRI), basically the origin and use of BOLD signal types used experiments. Further attention is paid fMRI data processing and statistical analysis. Subsequent chapters are devoted to a brief description of the most common software tools used to analyze data from fMRI. The main section was to create a program in MATLAB with a detailed graphic user interface for easy visualization and export output from analyzes of fMRI data. The second half is devoted to describing the program developer and graphic user interface, including key functionality. The final section describes the application program with real data from clinical studies of dynamic connectivity and use in an international project APGem.