京都大学 / 新制・課程博士 / 博士(医学) / 甲第23793号 / 医博第4839号 / 新制||医||1057(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 大森 孝一, 教授 花川 隆, 教授 渡邊 直樹 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 新制・課程博士 / 博士(人間健康科学) / 甲第23828号 / 人健博第99号 / 新制||人健||7(附属図書館) / 京都大学大学院医学研究科人間健康科学系専攻 / (主査)教授 椎名 毅, 教授 杉本 直三, 教授 辻川 明孝 / 学位規則第4条第1項該当 / Doctor of Human Health Sciences / Kyoto University / DFAM

Laser-based photo-thermal therapies can provide minimally-invasive treatment of cancers. Their effectiveness is limited by light penetration depth in tissue due to its highly scattering properties. The highly disordered refractive index distribution in tissue leads to multiple-scattering of incident light. It has been hypothesized that mechanical compression has a great potential to enhance the capabilities of laser therapy by inducing localized water transport, decreasing the refractive index mismatch, and decreasing the scattering coefficient of tissue. To better understand this process, we investigated the refractive index change of ex-vivo dog skin during mechanical compression using a swept-source optical coherence tomography (OCT) device built in our lab. The Lorentz-Lorenz rule of mixtures was applied to evaluate the water and protein weight fraction of tissue simultaneously. Results show that the refractive index of skin increased from 1.38 to 1.52 during compression and water content decreased about 60%-70% when the skin sample was compressed by 70%. In addition, we conducted compression experiments on human finger, palm, back of hand, and front of forearm in vivo. OCT images of these skin sites before and after compression by 1 minute were compared. Optical thickness of epidermis and light penetration depth in the dermis were measured. The extended Huygens-Fresnel model was applied to measure the scattering coefficient μs of skin specimens. μs of skin was measured to be about 10-17 mm-1 before compression and decreased 60%-80% after compression, which increases the averaged light intensity by 2-7 dB and almost doubles light penetration depth in dermis. It is quite significant in laser therapies especially for treating epithelia cancers which originate at 1-2 mm beneath the tissue surface. In the OCT imaging of skin dehydration experiment, we conclude that dehydration is an important mechanism of mechanical clearing. / Master of Science

Yes / Recent developments in optical coherence tomography (OCT) technology enable direct enface visualisation of retinal nerve fibre bundle (RNFB) loss in glaucoma. However, the optimum depth at which to visualise RNFBs across the retina is unknown. We aimed to evaluate the range of depths and optimum depth at which RNFBs can be visualised across the retina in healthy eyes. The central ± 25° retina of 10 healthy eyes from 10 people aged 57–75 years (median 68.5 years) were imaged with spectral domain OCT. Slab images of maximum axial resolution (4 μm) containing depth‐resolved attenuation coefficients were extracted from 0 to 193.5 μm below the inner limiting membrane (ILM). Bundle visibility within 10 regions of a superimposed grid was assessed subjectively by trained optometrists (n = 8), according to written instructions. Anterior and posterior limits of RNFB visibility and depth of best visibility were identified for each grid sector. Effects of retinal location and individual eye on RNFB visibility were explored using linear mixed modelling with likelihood ratio tests. Intraclass correlation coefficient (ICC) was used to measure overall agreement and repeatability of grading. Spearman’s correlation was used to measure correlation between depth range of visible RNFBs and retinal nerve fibre layer thickness (RNFLT). Retinal location and individual eye affected anterior limit of visibility (χ2(9) = 58.6 and 60.5, both p < 0.0001), but none of the differences exceeded instrument resolution, making anterior limit consistent across the retina and different eyes. Greater differences were observed in the posterior limit of visibility across retinal areas (χ2(9) = 1671.1, p < 0.0001) and different eyes (χ2(9) = 88.7, p < 0.0001). Optimal depth for visualisation of RNFBs was around 20 µm below the ILM in most regions. It varied slightly with retinal location (χ2(9) = 58.8, p < 0.0001), but it was not affected by individual eye (χ2(9) = 10.7, p = 0.29). RNFB visibility showed good agreement between graders (ICC 0.89, 95%CI 0.87–0.91), and excellent repeatability (ICC 0.96–0.99). Depth range of visible RNFBs was highly correlated with RNFLT (ρ = 0.9, 95%CI: 0.86–0.95). The range of depths with visible RNFBs varies markedly across the healthy retina, consistently with RNFLT. To extract all RNFB information consistently across the retina, slab properties should account for differences across retinal locations and between individual eyes. / This work was supported by a College of Optometrists Research Fellowship (JD).

Yes / Purpose: We present a subjective approach to detecting glaucomatous defects in enface images and assess its diagnostic performance. We also test the hypothesis that if reflectivity changes precede thickness changes in glaucoma there should be reduced correlation between the modalities in glaucoma compared to controls. Methods: Twenty glaucoma participants and 20 age-matched controls underwent high-resolution OCT scans of one eye. 4 μm-thick enface slabs were constructed through the retina. Enface indices were depths of first gap in visible retinal nerve fiber bundles (RNFBs) and last visible bundle, subjectively evaluated in six sectors of a 3.5 mm circle around the optic disc. Retinal nerve fiber layer thickness (RNFLT) along the same circle was extracted at angles corresponding to enface indices. Between-group differences were tested by linear mixed models. Diagnostic performance was measured by partial receiver operating characteristic area (pAUC). Results: First gap and last visible bundle were closer to the inner limiting membrane in glaucoma eyes (both P < 0.0001). Enface indices showed excellent diagnostic perfor mance (pAUCs 0.63–1.00), similar to RNFLT (pAUCs 0.63–0.95). Correlation between enface and RNFLT parameters was strong in healthy (r = 0.81–0.92) and glaucoma eyes (r = 0.73–0.80). Conclusions: This simple subjective method reliably identifies glaucomatous defects in enface images with diagnostic performance at least as good as existing thickness indices. Thickness and reflectivity were similarly related in healthy and glaucoma eyes, providing no strong evidence of reflectivity loss preceding thinning. Objective analyses may realize further potential of enface OCT images in glaucoma. Translational Relevance: Novel enface OCT indices may aid glaucoma diagnosis.

Este trabalho tem por objetivo propor um método eficiente para avaliar gestão de estoque, aplicando conjuntamente a Teoria de Controle Ótimo (OCT), para obter funções de estocagem dinamicamente ótimas, e Análise Envoltória de Dados (DEA), para calcular as eficiências relativas. Tendo em vista esse objetivo foi desenvolvido um modelo integrado DEA-OCT para calcular a eficiência de custo otimizada ao longo do tempo, quando o sistema possui variáveis relacionadas entre si, como no caso de sistemas de controle de estoque, e para analisar produção e demanda (assim como a variável estoque, oriunda dessa relação), estendendo o modelo variacional. Este trabalho aplica o modelo proposto a 647 empresas das Américas do Sul e do Norte, depois faz uma comparação entre Brasil e Chile (países emergentes economicamente), posteriormente focando no setor de comércio, considerando seus sistemas produção-estoque com dados de variáveis contábeis. Os modelos minimizam os custos de produção e de estoque para calcular a eficiência de custo ao longo do tempo. O output (produto, ou variável de saída) é a demanda; o input (insumo, ou variável de entrada) é a produção, e o intermediate (variável intermediária) é o estoque. Seus custos são considerados na função objetivo. É acrescentada uma restrição variacional da OCT para descrever a relação entre demanda, produção e estoque. Em resumo, o modelo é relevante por calcular eficiência prevenindo a possibilidade de obter uma projeção que ignora a relação entre as variáveis, uma vez que essa relação sempre ocorre, na prática, em sistemas de controle de estoque. As principais contribuições são: possibilitar o uso de OCT como a ferramenta de benchmarking DEA no contexto de eficiência dinâmica, estender o modelo DEA variacional de Sengupta (1995), incluindo restrições de modelos mais recentes e possibilitar o cálculo de eficiência quando há relação entre as variáveis. / This work aims to propose an efficient method to evaluate inventory management, jointly applying optimal control theory (OCT), obtaining dynamically optimal production and inventory functions, and data envelopment analysis (DEA), calculating the relative efficiencies. With this objective in mind, it was developed a DEA-OCT integrated model to calculate allocative efficiency optimized over time, when systems have variable with relationship among themselves, like in the case of inventory control systems, and for analyzing production and demand (as the inventory variable obtained from this relationship), extending the variational model. This paper applies the proposed model to 647 companies from South and North America, after that it was made a comparison between Brazil and Chile (economically emerging countries), then focusing on the commercial sector, considering its production-inventory systems and data from accounting variables. The model minimizes the inventory and production costs to calculate the allocative efficiency over time. The output is demand; the input is production, and the intermediate variable is inventory. Their costs are considered in the objective function. A variational constraint OCT is added to describe the relationship among demand, production, and inventory. In summary, the model is relevant to calculate efficiency by preventing the possibility of finding a projection that ignores the relationship among variables, since this relationship always occur in practice in inventory control systems. The main contributions are: using OCT as the benchmarking tool DEA in the context of dynamic efficiency, extending the Sengupta (1995) variational DEA model, including constraints from recent model and making it possible to calculate efficiency when there is a relationship among variables.

L’osteopetrosi autosomale recessiva (ARO) è un gruppo di malattie dovute a un difettoso funzionamento degli osteoclasti che preclude un rimodellamento osseo corretto. Nel 50% dei casi umani il difetto è dovuto ad una delezione nel gene Tcirg1. Il modello murino mutante oc/oc porta lo stesso difetto genetico e fenotipico umano. Nel lavoro di tesi si è dimostrato che gli epatociti fetali di 12.5 giorni di gestazione trapiantati in utero in feti mutati di 13.5 giorni di gestazione sono in grado di curare il fenotipo malato. Si è inoltre derivata una sottolinea di cellule staminali embrionali murine transgeniche per il costrutto plasmidico GOF18eGFP. Si vuole utilizzare la GFP sotto il controllo del promotore del gene Oct-4 come marcatore del livello di staminalità cellulare per microiniettare le ESC in blastocisti murine mutate oc/oc. / Autosomal recessive osteopetrosis (ARO) is a group of genetic disorders due to defects that preclude normal function of osteoclasts. In half the cases, human ARO is due to mutations in the Tcirg1 gene. The oc/oc mutant mouse closely recapitulates human Tcirg1-dependent ARO. In ths work we demonstrate that in utero injection of allogenic fetal liver cells on 12.5 days into oc/oc fetuses at 13.5 day post coitum completely rescue the osteopetrotic phenotype. Moreover, an embryonic stem cells line transgenic for GOF18eGFP was produced. The goal is to use the GFP under the transcriptional control of the Oct-4 promoter as a marker of pluripotency of the ESC that are to microinject into oc/oc blastocysts.

SWIFTS (Stationary Wave Integrated Fourier Transform Spectrometer) est une nouvelle technologie innovante de spectrométrie qui permet une réduction radicale de la taille des spectromètres à Transformée de Fourier, tout en conservant, et même en améliorant leurs performances. Grâce aux avancées de l'optique intégrée et des nanotechnologies, SWIFTS repose sur une méthode de détection optique originale, sans aucune partie mobile, où des nanoplots métalliques échantillonnent directement le champ évanescent d'une onde stationnaire dans un guide d'onde.Dans cette thèse, nous proposons de présenter le cheminement complet qui a mené, en partant du concept original, au développement puis à la mise en pratique de la technologie SWIFTS. Le document illustre notamment les caractérisations optiques, les choix technologiques et les optimisations entrepris pour la réalisation de spectromètres fonctionnels dans le domaine visible et proche-infrarouge. Des procédures de calibrages novatrices et complémentaires, basées sur du multiplexage fréquentiel et sur de l'interférométrie à faible cohérence temporelle, ont été développées pour déterminer avec précision les différentes irrégularités de fabrication et de comportement de l'appareil complètement intégré. Les spectromètres calibrés permettent à présent d'aborder des applications diverses en industrie et en recherche, de la caractérisation hautes performances de lasers, à l'interrogation de capteurs fibrés à réseaux de Bragg, aux techniques de spectrométries Raman et LIBS, et de tomographie optique OCT, jusqu'aux sciences de l'Univers (géophysique, astrophysique).SWIFTS est une innovation de rupture qui, de part sa miniaturisation obtenue sans compromis avec de hautes performances d'analyse spectrale, a la capacité de faire passer la spectrométrie du stade de la mesure complexe en laboratoire à celle d'un simple composant intégré pour des applications exigeantes. / SWIFTS (Stationary Wave Integrated Fourier Transform Spectrometer) is a new innovative technology of spectrometry that allows a drastic reduction of the size of Fourier transform spectrometers, while maintaining, and even improving their performance. With advances in integrated optics and nanotechnology, SWIFTS is based on an original method of optical detection, without any moving part, where metallic nanodots directly sample the evanescent field of a standing wave in a waveguide.In this thesis, we propose to present the complete process that led, starting from the original concept, to the development and the applications of the technology. The document illustrates the optical characterizations, the technological choices and the optimizations made for the realization of functional spectrometers in the visible and near-infrared range. Innovative and complementary procedures of calibrations, based on frequency multiplexing and low coherence interferometry, have been developed to accurately determine the various irregularities of the manufacturing and of the behavior of the integrated device. The calibrated spectrometers allow to address various applications in industry and research, such as high performance characterization of lasers, interrogation of fiber Bragg gratings sensors, Raman and LIBS spectrometry, optical coherence tomography OCT, and sciences of the Universe (geophysics, astrophysics).SWIFTS is a breakthrough innovation in spectrometry, without trade-off between miniaturization and high performance, that opens the way for product development based on the most demanding applications currently performed in research laboratories.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Selenium (Se) is an oligoelement crucial for various biological processes. Se has anti-inflammatory and antioxidant properties and plays a key role in brain development. Adenosine deaminase (ADA, EC 3.5.4.4) is a key enzyme in purine metabolism because it helps in the regulation of intracellular and extracellular levels of adenosine, an important nucleoside that acts in the neuromodulation of the immune and nervous systems. Oxidative stress occurs when there is an imbalance between the production of reactive species and their detoxification by systems that remove or repair the resulting damage. In this context, several Se compounds have been developed and studied, among them the 3-methyl-1-phenyl-2-(phenylseleno)oct-2-en-1-one (C12H2HOSe), which is an α, β-unsaturated ketone functionalized vinyl chalcogenide, and sodium selenate (Na2SeO4), the main inorganic form of Se found in animals and plants. The aim of this study was to compare the effects of C12H2HOSe (organic Se) and Na2SeO4 (inorganic Se) on the cell viability, lipoperoxidation and ADA activity in cortex slices of young rats. The results showed that only organic Se caused a reduction in ADA activity at the concentrations of 1, 10 and 30μM in the cerebral cortex slices of the rats tested. However, this result is not related to the oxidation of the thiol groups, since this compound did not alter the NP-SH and LDH levels. Both compounds did not affect the lipoperoxidation levels, although the organic Se was capable of sequestering NO radicals at the highest concentration tested. The inorganic Se protected the cerebral cortex slices against the sodium nitroprusside-induced damage and increased the NP-SH levels. The tests used to evaluate cell viability (LDH and MTT) suggest the maintenance of the cell integrity of the cortex slices exposed to Se compounds. Therefore, our results suggest that organic Se has immunomodulatory properties, due to the reduction in ADA activity, and acted in the maintenance of the cell integrity. In turn, the antioxidant activity of inorganic Se was reaffirmed. Hence, the results of this study paved the way to explore both Se compounds in the CNS. / O selênio (Se) é um oligoelemento essencial em vários processos biológicos. Possui propriedades antiinflamatórias, antioxidantes e desempenha um papel fundamental no desenvolvimento do cérebro. A adenosina deaminase (ADA, E.C. 3.5.4.4) é uma enzima chave no metabolismo das purinas, pois auxilia na regulação dos níveis intra e extracelulares de adenosina, um importante nucleosídeo que atua na neuromodulação dos sistemas nervoso e imune. O estresse oxidativo ocorre quando há desequilíbrio entre a produção de espécies reativas e sua desintoxicação através de sistemas que removem ou reparem os danos por elas causados. Nesse contexto, vários compostos de Se vêm sendo desenvolvidos e estudados, entre eles podemos citar o 3-metil-1-fenil-2-(fenilseleno)oct-2-en-1-um(C12H2HOSe),um organocalcogênio com uma cetona α, β- insaturada funcionando como um vinil calcogênio e o selenato de sódio (Na2SeO4), principal Se inorgânico encontrado em animais e plantas. Este estudo teve como objetivo comparar os efeitos do C12H2HOSe (Se orgânico) e Na2SeO4 (Se inorgânico) na viabilidade celular, parâmetros relacionados ao estresse oxidativo e atividade da ADA em fatias de córtex de ratos jovens. Os resultados demonstraram que apenas o Se orgânico provocou a diminuição na atividade da ADA nas concentrações de 1, 10 e 30μM nas fatias de córtex cerebral dos ratos testados. No entanto, esse resultado não tem relação com a oxidação de seus grupamentos tióis, já que esse composto não alterou os níveis de NP-SH e de LDH. Apesar de ambos os compostos não alteraram os níveis de lipoperoxidação, o Se orgânico apresentou capacidade de sequestrar radicais NO na maior concentração testada. Já o composto inorgânico de Se protegeu as fatias de córtex cerebral contra o dano induzido pelo nitroprussiato de sódio e aumentou os níveis de NP-SH. Os testes de viabilidade celular (LDH e MTT) sugerem a manutenção da integridade celular em fatias de córtex de ratos jovens quando expostos aos compostos de Se. Assim, nossos resultados sugerem que o Se orgânico apresenta propriedades imunomoduladoras por reduzir a atividade da ADA, além de atuar na manutenção da integridade celular. Já o Se inorgânico teve sua atividade antioxidante confirmada. Portanto, os resultados obtidos neste estudo destacam um caminho promissor a ser explorado por ambos os compostos no SNC.

Este trabalho tem por objetivo propor um método eficiente para avaliar gestão de estoque, aplicando conjuntamente a Teoria de Controle Ótimo (OCT), para obter funções de estocagem dinamicamente ótimas, e Análise Envoltória de Dados (DEA), para calcular as eficiências relativas. Tendo em vista esse objetivo foi desenvolvido um modelo integrado DEA-OCT para calcular a eficiência de custo otimizada ao longo do tempo, quando o sistema possui variáveis relacionadas entre si, como no caso de sistemas de controle de estoque, e para analisar produção e demanda (assim como a variável estoque, oriunda dessa relação), estendendo o modelo variacional. Este trabalho aplica o modelo proposto a 647 empresas das Américas do Sul e do Norte, depois faz uma comparação entre Brasil e Chile (países emergentes economicamente), posteriormente focando no setor de comércio, considerando seus sistemas produção-estoque com dados de variáveis contábeis. Os modelos minimizam os custos de produção e de estoque para calcular a eficiência de custo ao longo do tempo. O output (produto, ou variável de saída) é a demanda; o input (insumo, ou variável de entrada) é a produção, e o intermediate (variável intermediária) é o estoque. Seus custos são considerados na função objetivo. É acrescentada uma restrição variacional da OCT para descrever a relação entre demanda, produção e estoque. Em resumo, o modelo é relevante por calcular eficiência prevenindo a possibilidade de obter uma projeção que ignora a relação entre as variáveis, uma vez que essa relação sempre ocorre, na prática, em sistemas de controle de estoque. As principais contribuições são: possibilitar o uso de OCT como a ferramenta de benchmarking DEA no contexto de eficiência dinâmica, estender o modelo DEA variacional de Sengupta (1995), incluindo restrições de modelos mais recentes e possibilitar o cálculo de eficiência quando há relação entre as variáveis. / This work aims to propose an efficient method to evaluate inventory management, jointly applying optimal control theory (OCT), obtaining dynamically optimal production and inventory functions, and data envelopment analysis (DEA), calculating the relative efficiencies. With this objective in mind, it was developed a DEA-OCT integrated model to calculate allocative efficiency optimized over time, when systems have variable with relationship among themselves, like in the case of inventory control systems, and for analyzing production and demand (as the inventory variable obtained from this relationship), extending the variational model. This paper applies the proposed model to 647 companies from South and North America, after that it was made a comparison between Brazil and Chile (economically emerging countries), then focusing on the commercial sector, considering its production-inventory systems and data from accounting variables. The model minimizes the inventory and production costs to calculate the allocative efficiency over time. The output is demand; the input is production, and the intermediate variable is inventory. Their costs are considered in the objective function. A variational constraint OCT is added to describe the relationship among demand, production, and inventory. In summary, the model is relevant to calculate efficiency by preventing the possibility of finding a projection that ignores the relationship among variables, since this relationship always occur in practice in inventory control systems. The main contributions are: using OCT as the benchmarking tool DEA in the context of dynamic efficiency, extending the Sengupta (1995) variational DEA model, including constraints from recent model and making it possible to calculate efficiency when there is a relationship among variables.