Made available in DSpace on 2014-06-11T19:26:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-12-07Bitstream added on 2014-06-13T20:54:35Z : No. of bitstreams: 1 delmanto_a_me_botfm_prot.pdf: 1594635 bytes, checksum: 59b8850497a3a71e0f5ae2ec36c4ccb0 (MD5) / Fundação para o Desenvolvimento Médico e Hospitalar (Famesp) / Analisar o efeito do raloxifeno sobre o epitélio vaginal de mulheres pós-menopausa. Métodos: Estudaram-se prospectivamente entre novembro de 2004 a fevereiro de 2006, 80 mulheres na pós-menopausa. Quarenta pacientes receberam 6Omg/dia de raloxifeno (GR) e 40 mulheres compuseram o grupo não tratado (grupo controle, GC), pareado por idade e tempo de menopausa. O grupo tratado foi composto por pacientes com osteoporose de coluna lombar e/ou colo do fêmur. Foram excluídos aquelas com sinais e/ou sintomas de infecção do trato genital inferior e usuárias de terapia hormonal (TH) até seis meses prévios ao estudo. Os esfregaços vaginais foram coletados em dois momentos: inicial (MO) e após seis meses de seguimento (Ml). Para avaliação do epitélio vaginal foi utilizado o valor de maturação, com a contagem de células superficias, intermediárias e parabasais. Os esfregaços foram analisados por único citopatologista, sem conhecimento dos dados das pacientes. Para análise estatística empregou- se o teste t de Student, teste Wilcoxon Mann-Witney e o teste Qui-Quadrado. Resultados: Na comparação estatística inicial os grupos foram homogêneos. Comparando os momentos inicial e final, não foram observadas diferenças estatisticamente sígnífícativas nos valores medianos de maturação do epitélio vaginal e na porcentagem de células superficiais, intermediárias e parabasais entre os grupos. Não foi constatada correlação linear significativa entre o valor de maturação e a idade, o tempo de menopausa, o uso ou não de TH prévia, tabagismo e o índice de massa corpórea, em ambos os grupos. Conclusão: O tratamento com raloxifeno por seis meses não alterou o valor de maturação do epitélio vaginal em mulheres na pós-menopausa. / To analyze the effect of raloxifene on the vaginal epithelium of postmenopausal women. Methods: Eighty postmenopausal women were studied prospectively between November of 2004 and February of 2006. Forty patients received 6omglday of raloxifene (GR), and 40 women comprised the non-treated group (control group, CG), paired by age and time of menopause. The treated group was composed of patients with osteoporosis of the lumbar column and / or femur. Those with signs and / or symptoms of infection of the inferior genital tract and users of hormonal therapies (HT) up to six months prior to the study were excluded. Vaginal smears were collected at two moments: initial (MO) and after six months of follow-up (Ml). To evaluate the vaginal epithelium, the maturation value was determined, along with counts of superficial, intermediate and parabasal cells. Smears were analyzed by only one cytopathologist, without knowledge of patient data. For statistical analysis Student's t test, Wilcoxon Mann Witney test and Chi-Squared test were employed. Results: In the initial statistical comparison the groups were homogeneous. Comparing the initial and final moments, no statistically significant differences were observed in median values of vaginal epithelial maturation or in percentage of superficial, intermediate and parabasal cells between the groups. There was no significant linear correlation between value of vaginal epithelial maturation and age, time of menopause, use or not of previous HT, smoking or body mass index, in both groups. Conclusion: Treatment with raloxifene for six months did not alter the maturation value of vaginal epithelium in postmenopausal women.

Les cellules souches pluripotentes (CSP) incluent les cellules souches embryonnaires (ES) et les celles souches pluripotentes induites (iPS). Elles sont définies par deux propriétés fondamentales : l’auto-renouvellement et la capacité à se différencier dans tous les types cellulaires. Les ES sont dérivées de la masse cellulaire de l’embryon. Elles soulèvent l’intérêt de la communauté scientifique du fait de leur capacité à générer tous les tissus. Il s’agit d’un outil biotechnologique majeur dont les applications thérapeutiques et pharmacologiques comporteront notamment la médecine régénératrice, la modélisation in vitro de maladies humaines et le criblage de candidat-médicaments. Cependant l’utilisation d’embryons humains pour générer les ES soulève des problèmes éthiques. Les iPS contournent ces difficultés car elles sont dérivées de cellules somatiques différenciées. En effet, S. Yamanaka, qui a reçu le prix Nobel en 2012, a découvert en 2006 une technique simple de reprogrammation cellulaire. L’expression transitoire de quatre gènes (OCT4, SOX2, c-MYC and KLF4) est suffisante pour reprogrammer des fibroblastes murins en iPS. Ces cellules iPS ont la même morphologie et les mêmes propriétés que les cellules ES. L’année suivante, S. Yamanaka a appliqué avec succès son cocktail à des fibroblastes humains pour produire des iPS humaines (hiPS). Les hiPS peuvent également dépasser les problèmes immunologiques soulevés par l’utilisation d’ES dans la thérapie cellulaire, par le simple fait que les hiPS pourront être dérivées du patient à traiter. De plus, parce qu’il est possible de choisir les cellules du donneur à reprogrammer selon son génotype, il est plus facile de modéliser des maladies génétiques à partir d’hiPS que d’ES. Enfin, d’un point de vue pharmaceutique, les hiPS peuvent fournir une plateforme quasi-infinie pour le criblage de molécules afin de traiter diverses pathologies. Le but de mon projet de recherche est l’utilisation de la technologie hiPS afin de modéliser le développement de l’épithélium bronchique. Premièrement, in vivo, des tératomes ont été générés après injection d’hiPS dans des souris immunodéficientes. Les tératomes démontrent la capacité de nos hiPS à se différencier en épithélium bronchique. Secondairement, in vitro, reproduire le développement embryonnaire et fœtal permet d’offrir une méthode simple pour modéliser l’épithélium bronchique dans un puits. Cette technologie ouvre la voie vers de nombreuses recherches, du criblage de molécules à la production de cellules souches pour réparer l’épithélium bronchique, et in fine à la promotion de nouveaux traitements pharmacologiques ou de thérapie innovante pour les maladies respiratoires. / Pluripotent stem cells (PSC) include embryonic stem cells (ES) and induced pluripotent stem cells (iPS). They are defined by two fundamental properties: self-renewal and the capacity to differentiate into all cell types. ES cells are derived from the inner cell mass of embryos. They arouse the interest of the scientific community in particular for their ability to generate all tissues. They provide major therapeutic and pharmacological applications, including regenerative medicine, in vitro modelling of human diseases and molecular screening. However, the use of human blastocysts to generate ES cells raises many ethical problems. iPS circumvent these ethical issues as they can be derived from differentiated somatic tissues. Indeed, S. Yamanaka, Nobel Prize in 2012, discovered in 2006 a simple technique of cellular reprogramming. The transient expression of four genes (OCT4, SOX2, c-MYC and KLF4) is sufficient to reprogram mouse fibroblasts into iPS. These iPS cells have the same morphology and the same properties than ES cells. The following year, S. Yamanaka applied successfully his cocktail to human fibroblasts to produce human iPS (hiPS). hiPS may also overcome immunological problems raised by the use of ES cell for cellular therapy, as hiPS can be derived from the patient to be treated. In addition, it is easier to model genetic diseases from hiPS than ES, because it is possible to choose the donor cells to reprogram according to its genotype. Finally, from a pharmacological point of view, hiPS can provide a broad platform of molecular screening to treat various diseases. The aim of my research project is to use the hiPS technology to model the development of bronchial epithelium. First, in vivo, teratomas were formed by the injection of hiPS into immunodeficient mice. Teratomas highlight the ability of differentiation of our hiPS into bronchial epithelium. Second, in vitro, reproducing embryonic and foetal bronchial development provides a way to model bronchial epithelium in a dish.These techniques open the door to many potential research avenues from screening small molecules to engineering stem cells to repair bronchial epithelium, and will in fine promote new pharmacologic or cell-based treatments for respiratory diseases.

The products of the Cdx genes, Cdx1, Cdx2 and Cdx4, are known to play essential roles in many developmental processes including neural tube closure, axial elongation and patterning the anterior-posterior axis of the developing embryo. Cdx1 and Cdx2 are both expressed in the endoderm of the embryo and persist throughout adulthood in the intestinal epithelium, but their functions and mechanisms of action in this lineage are poorly understood, in part due to the peri-implantation lethality of Cdx2-/- mice. To circumvent this limitation, a conditional loss of function strategy was used to inactivate Cdx2 in the intestinal epithelium. These conditional mutants were also crossed to Cdx1-/- mice, which are viable and fertile, to examine potential functional compensation between these family members. The major findings of this study are that Cdx2 regulates patterning and differentiation of the small intestinal epithelium, while Cdx1 does not appear to make a contribution to either process. Furthermore, Cdx operates upstream of Notch ligand Delta-like 1 (Dll1) in endoderm and mesoderm derivatives, demonstrating that Cdx function is similar in different lineages. Finally, Cdx2 cannot fulfill the requirement for Cdx1 in regulation of its own promoter in the intestine. This is the first in vivo evidence that these two family members have context-dependent functional specificity. Altogether, this study underscores critical roles and mechanisms of action for Cdx members in the developing intestine and mesoderm.

The cellular mechanisms responsible for rectal acidification in the desert locust, Schistocerca gregaria, were investigated in isolated recta mounted as flat sheets in modified Ussing chambers. In the absence of exogenous CO₂, HCO₃⁻, and phosphate, the isolated rectum (under both open- and short-circuit current conditions) was capable of rates of net acid secretion (J[subscript]H+) similar to those observed in vivo, demonstrating the viability of the preparation and suggesting that rectal acidification was due to proton secretion rather than selective movements of HCO₃⁻ or phosphate. The possibility that trace levels of metabolic CO₂ might be generating sufficient HCO₃⁻ to account for the observed rates of rectal acidification (via HCO₃⁻ reabsorption) was assessed by adding exogenous CO₂/HCO₃⁻ to the contraluminal bath. The small increases in J[subscript]H+ observed after addition of 2% or 5% CO₂ were shown to be due to simple hydration of CO₂ which had diffused into the lumen (from the contraluminal bath), rather than changes in rates of HCO₃⁻ reabsorption. Since measurable quantities of luminal HCO₃⁻ did not directly affect the apical acid/base transport mechanism per se, it was concluded that metabolic CO₂ could not generate sufficient HCO₃⁻ in the lumen to account for the rates of rectal acidification observed under nominally CO₂/HCO₃⁻-free conditions and that J[subscript]H+ must be due to a proton secretory rather than bicarbonate reabsorptive mechanism. Microelectrode measurements of intracellular pH (pHi) and apical and basolateral membrane potentials (Va and Vb respectively) indicated that luminal pH was not in equilibrium with either contraluminal pH or pHi and that the mechanism responsible for active luminal acid secretion resided on the apical membrane. Preliminary measurements of bath total ammonia (ie. NH₃ + NH₄+) levels in the previous experiments suggested that the rectum was actively secreting ammonia at significant rates across the apical membrane into the lumen. If the ammonia crossed the apical membrane as NH₃ rather than NH₄+, rates of luminal ammonia secretion (J[subscript]Amm) would have to be added to J[subscript]H+ to obtain corrected values of luminal proton secretion. In the absence of exogenously added ammonia and CO₂, ammonia was preferentially secreted into the lumen under both open- and short-circuit current conditions. J[subscript]Amm was dependent on the presence of luminal amino acids and was relatively unaffected by K[superscript]+ removal or changes in luminal pH from 7.00 to 5.00. Bilateral Na+ substitution or luminal addition of ImM amiloride reduced J[subscript]Amm by 63% and 65% respectively. The data consistently demonstrate that the rectum secretes significant quantities of endogenously produced ammonia preferentially into the lumen as NH₄+ rather than NH₃ via an apical Na[superscript]+/NH₄[superscript]+ exchange mechanism. Clearly, rates of net acid secretion estimated by titratable acidity do not have to include a correction for luminal ammonia secretion. Although J[subscript]H+ was completely unaffected by changes in contraluminal pH, it could be progressively reduced (and eventually abolished) by imposition of either transepithelial pH gradients (lumen acid) or transepithelial electrical gradients (lumen positive). Under short-circuit current conditions, the bulk of J[subscript]H+ was not dependent on Na[superscript]+, K[superscript]+, CI⁻, Mg₂+, or Ca+ and was due to a primary electrogenic proton translocating mechanism located on the apical membrane. A small component (10-16%) of J[subscript]H+ measured under these conditions could be attributed to an apical amiloride-inhibitable Na[superscript]+/H[superscript]+ exchange mechanism. Inhibition of JH+ by anoxia or reduction of luminal pH unmasked a significant proton diffusional pathway on the apical membrane in parallel with the active proton pump. The fact that J[subscript]H+ was significantly inhibited (42%-66%) by contraluminal addition of ImM cAMP and relatively unaffected by changes in contraluminal pCO₂ or pH suggests that net acid secretion in the locust rectum in vivo is modulated by circulating hormonal factors rather than haemolymph pH or pCO₂ per se. / Science, Faculty of / Zoology, Department of / Graduate

京都大学 / 0048 / 新制・論文博士 / 博士(医学) / 乙第13265号 / 論医博第2179号 / 新制||医||1038(附属図書館) / (主査)教授 別所 和久, 教授 安達 泰治, 教授 戸口田 淳也 / 学位規則第4条第2項該当 / Doctor of Medical Science / Kyoto University / DFAM

Melatonin is an indolamine hormone presumably synthesized by retinal photoreceptors, and may act as a paracrine signal of darkness within the retina. Previous studies have suggested that melatonin, acting through specific receptors, may be involved in cyclic retinal functions such as photoreceptor outer segment disc shedding and phagocytosis, and modulation of neurotransmitter release in the inner retina. The goal of this study was to determine if melatonin receptor mRNA is expressed in the neural retina and retinal pigment epithelium (RPE) of Xenopus laevis. Sheets of RPE, devoid of contaminating cells, were obtained from Xenopus eyes, and epithelial cultures were subsequently established on microporous membrane filters in a defined medium. Total RNA was isolated from whole brain, neural retina, fresh RPE sheets, and cultured RPE cells. RNA expression of the three known Xenopus melatonin receptor subtypes (MEL1A, 1B, and 1C) was determined by reverse- transcription/polymerase chain reaction (RT/PCR) amplification, followed by Southern hybridization with RNA probes. PCR-amplified cDNA encoding melatonin receptor subtypes 1B and 1C, but not 1A, were detected in reverse-transcribed RNA obtained from brain, neural retina and RPE. RPE cells grown in culture for two weeks also demonstrated 1B and 1C receptor RNA expression. This study suggests that RNA encoding the 1B and 1C melatonin receptor subtypes is expressed in the neural retina and RPE of Xenopus retina, and the expression persists in RPE cells when grown in culture. The expression of melatonin receptor RNA in the RPE may reflect a regulatory role for melatonin in some diurnal events that occur in this tissue, such as phagocytosis of photoreceptor outer segment membranes, and intracellular migration of pigment granules.

No description available.

Members of the class Mammalia have the most advanced skeletal complexity of the nasal cavity among vertebrates. Most mammals have an olfactory epithelium that consists of basal cells, supporting cells and olfactory sensory neurons that bind odor molecules with their cilia. The olfactory epithelium is responsible for detecting odor stimuli. The surface area of olfactory epithelium varies greatly among species. Carnivores have a generally larger surface area of the olfactory epithelium than primates and ungulates of the same size. Modern odontocetes lack olfactory epithelium. To get an overview of the between-species differences of the olfactory epithelium surface area and number of olfactory receptor cells, a search of the scientific literature was performed, using the database Web of Science and references from the scientific articles. The assembled data were entered into two tables, one that contains species names, surface area of the olfactory epithelium and references, and another that includes the total number of olfactory receptor cells for the few species that have been studied in this respect so far. Methods of measuring olfactory epithelium size differ, some studies used immunohistochemistry, other measured osteological proportions, like the surface area of the olfactory turbinals. A compilation of the published data provides an overview of the range that the size of the olfactory epithelium can have and allows for between-species comparisons of this anatomical measure as well as for assessing possible correlations with olfactory capabilities.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20804号 / 医博第4304号 / 新制||医||1025(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 河本 宏, 教授 松田 文彦, 教授 柳田 素子 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

No description available.