Age related macular degeneration (AMD) is one of the leading causes of blindness amongst the elderly. Macular pigment (MP) in the retina has been established to protect individuals against AMD. Improving levels of MP by diet or supplements is the constant quest of clinical practitioners and researchers, thus necessitating development of instruments capable of repeatable and reliable MP measurement. Cataract, a consequence of the rising opacity levels of the lens with age is one of the other major causes of blindness in the world. Mapcatsf, a LED-based microprocessor-controlled heterochromatic flicker photometer (HFP) using photopic vision is capable of measuring the levels of MP and the opacity of the lens in terms of optical density. Test-retest measurements conducted on 83 subjects were analyzed for repeatability in macular pigment optical density (MPOD) measurements. Reliability of the lens optical density (LOD) measurements were tested and compared with those obtained at absolute scotopic thresholds for 25 individuals. A supplement study with 32 individuals both in the young (50) age groups for 6 months further established Mapcatsf’s capacity to monitor changing levels of MP in individuals. As an overall outcome, high levels of repeatability and reliability were obtained in MPOD and LOD measurements establishing Mapcatsf as an instrument for use in clinical settings in the future.

Diversos procedimentos endodônticos dependem da utilização das radiografias periapicais. O exame convencional ainda é o método de eleição para a prática clínica, a despeito dos avanços trazidos com os sistemas digitais e com os localizadores eletrônicos. Ocorre, porém, que nos estudos científicos não se verifica adequada padronização quanto aos modelos de estudo utilizados. Ao mensurar e comparar a densidade óptica das imagens do canal e dentina radicular de radiografias periapicais de um modelo experimental e de indivíduos vivos, este trabalho se propôs avaliar a viabilidade da utilização deste modelo em estudos endodônticos radiográficos. Após coleta de dados avalizada cientificamente pela utilização do aparelho fotodensitômetro (MRA), e análise estatística (Testes de Kolmogorov-Smirnof, Levene com correção de Bonferroni, Kruskal-Wallis, Mann-Whitney e de correlação de Pearson), observou-se que os resultados obtidos não evidenciaram diferenças estatisticamente significativas entre as radiografias do modelo e dos pacientes. Diante disso, conclui-se que o modelo experimental é adequado para a utilização em experimentos endodôntico-radiográficos. / Several endodontic procedures depend of dental radiographs. The conventional radiograph is the usual method to clinical practice, even the advances proportionate by digital systems and electronic apex locators. However, the endodontic studies dont display standardized radiographic models. Analyzing the optical density of the images from an experimental model and patients, based on digital photodensitometer (MRA), this study evaluated the viability of this model in endodontic studies. The Kolmogorov-Smirnof, Levene, Bonferronis correction, Kruskal-Wallis, Mann-Whitney and Pearsons correlation tests dont display statistical differences between images from patients and the model. Thus concluded that is adequate to use in radiographicendodontic studies.

<p> Some experiments designed to test the validity of the edge-spread function (ESF) model for neutron radiographic image formation are described; in addition the experiments are meant to illustrate the application of ESF methods to two areas of practical concern. First, the prediction of optical density curves for specified material and geometric configurations is considered; then, the use of ESF methods in dimensioning irradiated reactor fuel elements is examined. Overall, the results indicate that within the framework of assumptions which ESF theory is based upon, the correlation between theory and experiment is excellent. The results also suggest that in situations which deviate from the theoretical ideal, the ESF method may serve as a good first approximation to more complex models. </p> / Thesis / Master of Engineering (ME)

There is significant interest by the food industry in applying milder processing conditions. A major area of research within predictive modelling has been the search for models which accurately predict the effect of combining multiple processes or hurdles. For a mild process, which has temperature as the major microbial injury step, the effect of the other combined hurdles in inhibiting growth of the injured organisms must be understood. The latter means that the inoculum size dependency of the time to growth must also be fully understood. This essentially links injury steps with the potential for growth. Herein, we have been developing the use of optical density (O.D) for obtaining growth rates and lag times using multiple inocula rather than using the traditional methods which use one single inoculum. All analyses were performed in the Bioscreen analyser which measures O.D. The time to detection (TTD) was defined as the time needed for each inoculum to reach an O.D=0.2 and O.D was related to microbial numbers with simple calibration curves. Several primary models were used to predict growth curves from O.D data and it was shown that the classic logistic, the Baranyi and the 3-phase linear model (3-PLM) were the most capable primary models of those examined while the modified Gompertz and modified logistic could not reproduce TTD data. Using the Malthusian approximation of the logistic model the effect of mild temperature shifts was studied. The data obtained showed that for mild temperature shifts, growth rates quickly changed to the new environment without the induction of lags. The growth of Listeria monocytogenes, Salmonella Typhimurium and Escherichia coli was studied at 30⁰C and/or 37⁰C, in different NaCl concentrations, pH and their combinations. The classical 3-parameter logistic with lag model was rearranged to provide the theoretical foundation for the observed TTD and accurate growth rates and lag times could be estimated. As the conditions became more unfavourable, the lag time increased while the growth rate decreased. Also, the growth rate was found to be independent from the inoculum size; the inoculum size affected only the TTD. The Minimum Inhibitory Concentration (MICNaCl and MICpH) was calculated using the Lambert and Pearson model (LPM) and also the Growth/No Growth (G/NG) interface was determined using combinations of NaCl and pH. These data were transformed in rate to detection (RTD) and fitted with a response surface model (RSM) which was subsequently compared with the Extended LPM (ELPM). The LPM and the ELPM could analyse results from individual and combined inhibitors, respectively. Following a mild thermal process a lag due to thermal injury was also induced, the magnitude of which was dependent on the organism and environmental conditions; the observed distribution of the lags appeared, in general, to follow the Log-normal distribution. After the lag period due to injury, growth recommenced at the rate dictated by the growth environment present. Traditional growth curves were constructed and compared with the data obtained from the Bioscreen under the same conditions. From the results obtained, it can be suggested that the increased lag times and growth rates obtained from the traditional plate counts compared with the values obtained from the Bioscreen microbiological analyser, might be an artifact of the plating method or may be due to the use of the modified Gompertz to study the growth. In conclusion, O.D can be used to accurately determine growth parameters, to give a better understanding and quantify the G/NG interface and to examine a wealth of phenomena such as fluctuating temperatures and mild thermal treatments. The comparison between the traditional growth curves against the data obtained from the Bioscreen showed that the TTD method is a rapid, more accurate and cheaper method than the traditional plate count method which in combination with the models developed herein can offer new possibilities both to the research and the food industry.

Work size - 60 pages, including 35 pictures, 1 table. List of literature - 44 sources. The beginning of the work -2004 09 01, the end of the work - 2006 05 15. Purpose of work: To explore, what influence the additive lysozyme on technological properties of milk which are important in manufacture of fermental cheeses and sour - milk products has. In work presents the analysis of lysozyme influence on the technological properties of. The results show that lysozyme prevent to develop of undesirable microorganisms and positively influences on the quality of fermented milks. It was established that the additive of lysozyme prolongs the duration of the bactericidal phase. The investigation of the rennet formation time has shown that the clothing of the milk in samples with lysozyme formed 12 - 15  faster as in compared with the control sample without lysozyme. Besides, it is established, that the additive of lysozyme intensifies the process removal of the whey. The research investigation show that in samples with lysozyme, whey distinguish in smaller optical density as compared with control samples. The development of lactic bacteria during fermentation process was examined too. It was found that lysozyme influence on this process is very insignificant. It was established that the additive of lysozyme insignificant reduces viscosity and acidity of fermented milk gels.

It was investigated the colour reproduction on prints made in ink-jet printer. Experimentally investigated gradation characteristics of CMYK colours. It was made a 26 step strip with raster dots from 0 to 100 %. Prints were made at different printing regimes on papers with different whiteness. Zone optical densities were measured, compared with theoretical density values and gradational distortions were determined. Also were measured 100% raster strips in CIE L*a*b* coordinate system and defined the maximum reproduction of colours in ink-jet printer.

Made available in DSpace on 2014-12-17T15:37:15Z (GMT). No. of bitstreams: 1 AlaneMS_DISSERT1red.pdf: 4031089 bytes, checksum: da896508e54d2cd0138d081ad42b95de (MD5) Previous issue date: 2013-03-27 / Funda??o de Apoio ? Pesquisa do Estado do Rio Grande do Norte / The midline/intralaminar nuclei form a remarkable group of nuclei of the medial and dorsal thalamus. The midline nuclei, in rats, comprises the paratenial nuclei (PT), paraventricular (PV), intermediodorsal (IMD), reuniens (Re) and rhomboid (Rh). The intralaminar nuclei comprises the central medial (CM), paracentral (PC), central lateral (CL) and parafascicular (PF). Such nuclei have dense serotonergic innervation originating from the brainstem, especially from the so-called ascending activation system. These nuclei, in turn, send projections to various cortical and subcortical areas, specifically to limbic areas, which suggests the important role of this neurotransmitter in the limbic circuitry. The aim of this study was to characterize the distribution pattern and morphology of serotonin fibers in the nuclei of the midline and intralaminar thalamic of rocky cavy (Kerodon rupestris), a tipical rodent from brazilizan northeast. To reach this aim we used four rock cavies adults. Following the transcardially perfusion with paraformaldehyde and brain microtomy steps was performed immunohistochemistry for serotonin (5-HT), Nissl technique and subsequent achievement and image analysis to characterize the cytoarchitecture of these nuclei and the serotonergic fibers visualized. An analysis was made of Relative Optical Density (ROD) to semi-quantify the concentration of serotonin fibers in the areas of interest. Thus, we observed a cytoarchitectonic arrangement of these nuclei similar to that found in rats. In case of fibers distribution, those immunoreactive to 5-HT were presented in a higher concentration according as ROD in the midline nuclei relative to intralaminar; Re being the core which has a higher pixel value followed by the PV , Rh, IMD and PT. In intralaminar CL showed higher pixels, followed by nuclei CM, PC and PF. The serotonergic fibers were classified as number of varicosities and axon diameter, therefore find three types of fibers distributed through this nuclear complex: fibers rugous, granular and semi-granular. In PV fibers predominated rugous; in PT fibers predominated granular; IMD, CL and PF fibers were represented by semi-granular and Re, Rh, PC and CM fibers showed granular and semi-granular. Morphological characterization of serotonergic fibers and differences in density between the nuclei may suggest different patterns of synaptic organization of this neurotransmitter beyond confirming his large repertoire functional / O complexo nuclear da linha m?dia/intralaminar forma um not?vel grupo de n?cleos da parte medial e dorsal do t?lamo. Os n?cleos da linha m?dia, em ratos, s?o compreendidos pelos n?cleos paratenial (PT), paraventricular (PV), intermediodorsal (IMD), reuniens (Re) e romb?ide (Rh). J? os intralaminares compreendem os n?cleos central medial (CM), paracentral (PC), central lateral (CL) e parafascicular (PF). Tais n?cleos apresentam densa inerva??o seroton?rgica oriunda do tronco encef?lico, a partir, principalmente, do denominado sistema de ativa??o ascendente. Esses n?cleos, por sua vez, emitem proje??es para diversas ?reas corticais e subcorticais, mais especificamente para ?reas l?mbicas, o que prop?e o importante papel desse neurotransmissor na circuitaria l?mbica. O objetivo desse trabalho foi caracterizar o padr?o de distribui??o e morfologia das fibras de seroton?rgicas nos n?cleos da linha m?dia e intralaminares do t?lamo do moc? (Kerodon rupestris), um roedor t?pico da regi?o Nordeste. Para isso, foram utilizados 4 moc?s adultos jovens. Ap?s etapas de perfus?o e microtomia, foi realizada imunoistoqu?mica para serotonina (5-HT), t?cnica de Nissl e posterior coleta e an?lise de imagens a fim de caracterizar a citoarquitetura desses n?cleos, bem como as fibras de 5-HT neles visualizadas. Foi realizada ainda uma an?lise de Densidade ?ptica Relativa (DOR) para semiquantificar a concentra??o de fibras de 5-HT nas ?reas de interesse. Sendo assim, observamos uma distribui??o citoarquitet?nica desses n?cleos semelhante ao observado em ratos. Em se tratando da distribui??o de fibras, aquelas imunorreativas a 5-HT apresentaram-se em maior concentra??o, conforme a DOR, nos n?cleos da linha m?dia em rela??o aos intralaminares, sendo o Re o n?cleo que apresenta maior valor de pixels, seguido do PV, Rh, IMD e PT. Nos intralaminares o CL apresentou maior valor de pixels, seguido dos n?cleos CM, PC e PF. As fibras seroton?rgicas foram classificadas conforme n?mero de varicosidades, bem como di?metro axonal. Assim, encontramos tr?s tipos de fibras distribu?das atrav?s desse complexo nuclear: fibras rugosas, granulares e semi-granulares. No PV predominaram fibras rugosas; PT predominaram fibras granulares; IMD, CL e PF foram representados por fibras semi-granulares e Re, Rh, PC e CM apresentaram fibras granulares e semi-granulares. A caracteriza??o morfol?gica das fibras seroton?rgicas encontradas e as diferen?as de densidades entre os n?cleos permite sugerir diferentes padr?es de organiza??o sin?ptica deste neurotransmissor al?m de confirmar seu grande repert?rio funcional

Une activité germinale persiste après la naissance dans des niches spécialisées du cerveau des mammifères, à savoir le gyrus denté de l'hippocampe et la zone sous-ventriculaire (SVZ) bordant le ventricule latéral. Les cellules souches neurales (NSC) de la SVZ postnatale se différencient en progéniteurs transitoires qui vont générer des neuroblastes migrant à travers la voie de migration rostrale vers le bulbe olfactif, où ils se différencient en neurones. La SVZ génère également des progéniteurs gliaux qui se dispersent dans le parenchyme voisin. Les travaux récents auxquels j'ai participé soulignent la nature hétérogène de la SVZ postnatale, composée de différents microdomaines générant des lignées neurales distinctes. Les objectifs de mon travail de thèse ont permis de : 1) développer de nouveaux moyens pour explorer l'hétérogénéité de la SVZ; et 2) d'identifier et d'étudier le rôle d'un facteur de transcription exprimé par une sous population des NSCs de la SVZ. Objectif 1: La SVZ est une région hautement complexe et irrégulière dans laquelle une forte activité germinale persiste après la naissance. Le caractère hétérogène de la SVZ est évident et des études récentes ont généré une très grande base de données de transcrits, qui sont différentiellement exprimés entre les microdomaines. Cependant, un outil approprié pour l'analyse rapide du niveau d'expression d'une protéine d'intérêt, le long des axes rostro-caudal et dorso-ventral de la SVZ est toujours manquant et nécessaire. Par conséquent, j'ai développé "FlashMap", un logiciel semi-automatique qui permet une analyse rapide des niveaux d'expression de protéines dans le SVZ, basé sur des mesures de densité optique après immunohistochimie. "FlashMap" génère des cartes thermiques facilement lisibles en deux dimensions, qui peuvent être superposées avec précision aux reconstructions tridimensionnelles du système ventriculaire pour une visualisation spatiale fine et rapide. Cette nouvelle approche accélérera la recherche sur la régionalisation de la SVZ, en permettant l'identification de marqueurs (e.g. facteurs de transcription) exprimés dans des régions discrètes de la SVZ. Objectif 2: J'ai utilisé des approches de transcriptomique et de « fate mapping » des NSCs pour étudier la relation entre l'expression régionale de facteurs de transcription et leur différenciation dans des lignées neurales distinctes. Mes résultats supportent un amorçage précoce des NSCs à produire différents types cellulaires en fonction de leur localisation spatiale dans la SVZ. Nos données identifient Hopx comme un marqueur d'une sous population de NSCs qui génère principalement des astrocytes. De façon intéressante, la manipulation de l'expression de Hopx montre des effets mineurs sur l'astrogénèse, mais entraîne des changements marqués quant au nombre de NSCs et de leur descendance. Dans son ensemble, Mes résultats mettent en évidence à la fois une hétérogénéité spatiale des NSCs postnatales ainsi que leur amorçage précoce à produire des types cellulaires distincts / Germinal activity persists in the postnatal mammalian brain in specialized niches, namely the dentate gyrus of the hippocampus and the subventricular zone (SVZ) surrounding the lateral ventricle. Neural stem cells (NSCs) of the postnatal SVZ differentiate into transient amplifying progenitors that will generate neuroblasts migrating through the rostral migratory stream, into the olfactory bulb, where they differentiate into neurons. The SVZ additionally generates glial progenitors that invade the nearby parenchyma. Recent work to which I have participated, highlights the heterogeneous nature of the postnatal SVZ in respect to different microdomains generating distinct neural lineages. The objectives of my PhD work were twice: 1) to develop new means to explore the heterogeneity of the SVZ; and 2) to identify transcription factors expressed by subpopulations of NSCs of the SVZ and acting in their differential specification. Objective 1: The SVZ is a highly complex and irregular region of ongoing postnatal germinal activity. The heterogeneous character of the SVZ is evident and recent studies generated enormous datasets of transcripts, which are differentially expressed between divergent microdomains. However, an appropriate tool for fast analysis of the protein level along the full rostro-caudal and dorso-ventral extend of the SVZ is still missing. Therefore, I developed “FlashMap”, a semi-automatic software that allows rapid analysis of protein levels in the full SVZ, based on optical density measurements after immunohistochemistry. “FlashMap” generates easy readable heatmaps in two dimensions, which can be accurately superimposed on three-dimensional reconstructions of the ventricular system for rapid spatial visualization and analysis. This new approach will fasten research onto SVZ regionalization, by guiding the identification of markers, such as transcription factors expressed in specific SVZ microdomains. Objective 2: I used transcriptomic as well as fate mapping approaches to investigate the relation between regional expression of transcription factors by NSCs and their acquisition of distinct neural lineage fates. Our results support an early priming of NSCs to produce defined cell types depending of their spatial location in the SVZ and identify Hopx as a marker of a subpopulation biased to generate astrocytes. Interestingly, manipulation of Hopx expression showed minor effects on astrogenesis, but resulted in marked changes in the number of NSCs and of their progenies. Taken together, our results highlight transcriptional and spatial heterogeneity of postnatal NSCs, as well as their early priming toward specific lineages and suggest a role for Hopx in the evolution of SVZ germinal activity

La adaptación de la levadura Saccharomyces cerevisiae a condiciones de alta osmolaridad está mediada por la vía de HOG ((high-osmolarity glycerol). La activación de esta vía induce una serie de respuestas que van a permitir la supervivencia celular en respuesta a estrés. La regulación génica constituye una respuesta clave para dicha supervivencia. Se han descrito cinco factores de transcripción regulados por Hog1 en respuesta a estrés osmótico. Sin embargo, éstos no pueden explicar la totalidad de los genes regulados por la MAPK Hog1. En el presente trabajo describimos cómo el complejo transcripcional formado por las proteínas Rtg1 y Rtg3 regula, a través de la quinasa Hog1, la expresión de un conjunto específico de genes. Hog1 fosforila Rtg1 y Rtg3, aunque ninguna de estas fosforilaciones son esenciales para regulación transcripcional en respuesta a estrés. Este trabajo también muestra cómo la deleción de proteínas RTG provoca osmosensibilidad celular, lo que indica que la integridad de la vía de RTG es esencial para la supervivencia celular frente a un estrés osmótico. / In Saccharomyces cerevisiae the adaptation to high osmolarity is mediated by the HOG (high-osmolarity glycerol) pathway, which elicits different cellular responses required for cell survival upon osmostress. Regulation of gene expression is a major adaptative response required for cell survival in response to osmotic stress. At least five transcription factors have been reported to be controlled by the Hog1 MAPK. However, they cannot account for the regulation of all of the genes under the control of the Hog1 MAPK. Here we show that the Rtg1/3 transcriptional complex regulates the expression of specific genes upon osmostress in a Hog1-dependent manner. Hog1 phosphorylates both Rtg1 and Rtg3 proteins. However, none of these phosphorylations are essential for the transcriptional regulation upon osmostress. Here we also show that the deletion of RTG proteins leads to osmosensitivity at high osmolarity, suggesting that the RTG-pathway integrity is essential for cell survival upon stress.

L'activació sostinguda de Hog1 porta a una inhibició del creixement cel·lular. En aquest treball, hem observat que el fenotip de letalitat causat per l'activació sostinguda de Hog1 és parcialment inhibida per la mutació del complexe SCFCDC4. La inhibició de la mort causada per l'activació sostinguda de Hog1 depèn de la via d'extensió de la vida. Quan Hog1 s'activa de manera sostinguda, la mutació al complexe SCFCDC4 fa que augmenti l'expressió gènica depenent de Msn2 i Msn4 que condueix a una sobreexpressió del gen PNC1 i a una hiperactivació de la deacetilassa Sir2. La hiperactivació de Sir2 és capaç d'inhibir la mort causada per l'activació sostinguda de Hog1. També hem observat que la mort cel·lular causada per l'activació sostinguda de Hog1 és deguda a una inducció d'apoptosi. L'apoptosi induïda per Hog1 és inhibida per la mutació al complexe SCFCDC4. Per tant, la via d'extensió de la vida és capaç de prevenir l'apoptosi a través d'un mecanisme desconegut. / Sustained Hog1 activation leads to an inhibition of cell growth. In this work, we have observed that the lethal phenotype caused by sustained Hog1 activation is prevented by SCFCDC4 mutants. The prevention of Hog1-induced cell death by SCFCDC4 mutation depends on the lifespan extension pathway. Upon sustained Hog1 activation, SCFCDC4 mutation increases Msn2 and Msn4 dependent gene expression that leads to a PNC1 overexpression and a Sir2 deacetylase hyperactivation. Then, hyperactivation of Sir2 is able to prevent cell death caused by sustained Hog1 activation. We have also observed that cell death upon sustained Hog1 activation is due to an induction of apoptosis. The apoptosis induced by Hog1 is decreased by SCFCDC4 mutation. Therefore, lifespan extension pathway is able to prevent apoptosis by an unknown mechanism.