Orientador: Valéria Helena Alves Cagnon Quitete / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-23T04:47:08Z (GMT). No. of bitstreams: 1 Kido_LarissaAkemi_M.pdf: 14026607 bytes, checksum: 4b5dcdcd04b3cf312f7b62cb4cf2ac2b (MD5) Previous issue date: 2013 / Resumo: A senescência está associada a mudanças significativas no ambiente hormonal, sendo fator causador de alterações morfofuncionais na próstata. Os diferentes processos biológicos que ocorrem na próstata são regulados por polipeptídeos, dentre esses os fatores de crescimento do endotélio vascular (VEGF) e Endostatina, relacionados à angiogênese. Além disso, inibidores da enzima 5_ redutase-II, como a finasterida, tem papel importante no combate às doenças prostáticas. Assim, os principais objetivos desse estudo foram avaliar os efeitos estruturais e moleculares das terapias antiangiogênicas e da finasterida sobre a próstata ventral de camundongos durante a senescência. Noventa camundongos machos FVB de 18 e 52 semanas de idade foram divididos nos seguintes grupos: Jovem (JV) e Senil (SEN), os quais receberam injeções de Solução Fisiológica 0,9% (5 mL/Kg/dia s.c.); Finasterida (FIN): injeções de Finasterida (20 mg/Kg, s.c.); SU5416 (SU): SU5416 (6 mg/Kg, i.p.); TNP-470 (TNP): injeções de TNP-470 (15 mg/Kg, s.c.), e SU5416 + TNP-470 (SU+TNP): os mesmos tratamentos dos grupos SU e TNP. Após 21 dias de tratamento, amostras do lobo ventral da próstata foram coletadas e submetidas às análises morfológicas, imunohistoquímicas e Western Blotting. Os resultados demonstraram alterações moleculares e estruturais no microambiente prostático durante a senescência, como atrofia presença de células inflamatórias, e lesões proliferativas, as quais foram interrompidas e ou bloqueadas através dos tratamentos com as drogas antiangiogênicas e pela finasterida. Os resultados moleculares revelaram no grupo senil a diminuição das reatividades para AR e Endostatina, e aumento para ER-_, ER-_ e VEGF, quando comparados aos camundongos jovens. Os camundongos dos grupos tratados com finasterida, SU5416 e SU5416+TNP-470, quando comparados aos do grupo senil, demonstraram de forma geral diminuição das reatividades de VEGF e ER-_ e aumento de ER-_. Já o tratamento com TNP-470 foi marcado principalmente pela redução da reatividade e dos níveis protéicos de AR e ER-_, quando comparado aos grupos jovem e senil. Desta maneira, conclui-se que a senescência favoreceu a ocorrência de alterações estruturais e/ ou funcionais que sugerem o aparecimento de lesões malignas, em virtude do desequilíbrio na sinalização entre epitélio e estroma. O tratamento com finasterida, SU5416 e SU5416+TNP- 470 mostraram-se mais ativos na regulação dos processos proliferativos através da via estrogênica / Abstract: Senescence is associated with significant changes in the hormonal environment and is a cause of morphological and functional changes in the prostate. The different biological processes that occur in the prostate are influenced by different factors such as vascular endothelial growth factor (VEGF) and Endostatin, related to angiogenesis. Also, 5_-reductase inhibitors, such as finasteride, play an important role in treatment of prostatic diseases. Thus, the aims of this study were to evaluate the structural and molecular effects of antiangiogenic therapies and finasteride on the ventral prostate of mice during senescence. Ninety 52 and 18 week old male FVB mice, were divided into groups: Young (YNG) and Senile (SEN) groups, which received 0.9% saline (5 mL/kg/day sc) injections; Finasteride (FIN) group: Finasteride (20 mg/kg, sc); SU5416 (SU) group: SU5416 (6 mg/kg, ip) injections; TNP-470 (TNP) group: TNP-470 (15 mg/kg, sc) injections and SU5416+TNP-470 (SU+TNP470) group: The same treatment as the SU and TNP-470 groups. After 21 days of treatment, samples of the ventral lobe of the prostate were collected and analyzed for morphological, immunohistochemical and Western Blotting analyses. The results demonstrated structural and molecular changes in the prostatic microenvironment during senescence, such as atrophy, inflammatory cells, and proliferative lesions, which were interrupted and/or blocked by treatment with antiangiogenic drugs and finasteride. The molecular results revealed decreased reactivity for AR and Endostatin, and an increase for ER-_, ER-_ and VEGF in the senile group, when compared to young mice. The mice in the groups treated with finasteride, SU5416 and SU5416 + TNP-470, when compared to the senile group, showed in general decreased VEGF and ER-_ reactivities and increased ER-_ reactivity. The treatment with TNP-470 however, was marked mainly by reduced AR and ER-_ reactivity and protein levels, when compared to young and senile groups. Thus, it can be concluded that senescence contributed to the occurrence of structural and/or molecular alterations that suggest the onset of malignant lesions, due to the imbalance in the signaling between the epithelium and stroma. Treatments with finasteride, SU5416 and SU5416+TNP-470, were active in the regulation of proliferative processes by means of the estrogen pathways / Mestrado / Anatomia / Mestra em Biologia Celular e Estrutural

Orientador: Karina Gottardello Zecchin / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-19T22:07:17Z (GMT). No. of bitstreams: 1 Gomes_CamillaBorgesFerreira_M.pdf: 2172456 bytes, checksum: 5dd9bb49385077ce4213db57e518b145 (MD5) Previous issue date: 2012 / Resumo: O comprometimento ósseo em pacientes com câncer é responsável por significativas morbidade e redução da qualidade de vida, principalmente quando se trata de câncer de mama, no qual o esqueleto é o principal sítio de desenvolvimento das metástases. Bisfosfonatos (BPs) contendo ou não nitrogênio são os principais agentes usados no combate à perda óssea como na osteoporose, uma vez que induzem apoptose de osteoclastos. Recentemente foram demonstradas ações antitumoral e antiangiogência dos BPs, in vitro e in vivo. Desse modo, o objetivo deste trabalho foi analisar e comparar os efeitos antitumorais e antiangiogênicos de BP que não contêm nitrogênio (clodronato, CLO) e BP contendo nitrogênio (ácido zoledrônico, ZOL), em linhagem celular derivada de câncer de mama, MCF-7, e em culturas de células endoteliais. Os resultados mostram que ZOL reduziu a proliferação das MCF-7 de maneira dose-dependente, com aumento do número de células nas fases G0/G1, sem alterar a viabilidade. Paralemamente, CLO não mostrou nenhum efeito sobre as células tumorais. O tratamento de células endoteliais de coelho, RAEC, com ZOL ou CLO reduziu a proliferação celular derivada, sem alterar sua viabilidade. Células endoteliais humanas HUVEC mostraram 15% de redução da viabilidade quando tratadas com ZOL, enquanto CLO não mostrou efeitos citotóxicos. A incubação de células HUVEC com meio condicionado por células MCF-7 tratadas com ZOL culminou na redução da proliferação das células endoteliais e na redução da formação de estruturas semelhantes a vasos, in vitro. Esse mesmo meio condicionado pelas células tumorais tratadas com ZOL resultou em menor proliferação de células provenientes de aorta murina, em ensaio ex vivo. Ensaios de imunoabsorbância revelaram diminuição nos níveis de VEGFA total nesse meio condicionado, assim como aumento da quantidade da isoforma antiangiogênica VEGFA165b, após tratamento das MCF-7 com ZOL. Por último, o tratamento das células MCF-7 com ZOL modulou a expressão de VEGFA total e de suas isoformas. Em conjunto, estes achados mostram que ZOL exibe ação antitumoral e antiangiogênica em células de câncer de mama e em células endoteliais. Tais dados contribuem para um maior esclarecimento sobre os mecanismos de ação dos BPs, permitindo buscas por melhorias no uso desses agentes como coadjuvantes na quimioterapia, a fim de minimizar a morbidade decorrente de metástases ósseas em tumores malignos / Abstract: Patients with cancer frequently develop bone metastasis, resulting in considerable morbidity and affecting quality of life. This is particularly important considering breast cancer, in which skeletal is the main site for metastasis. Non-nitrogen-containing and nitrogen-containing bisphosphonates (BPs) are the mainly agents to treat bone loss due to osteoporosis, Paget disease, bone metastasis, multiple myeloma, hypercalcemia and osteogenesis imperfecta. BPs impair bone loss by inducing apoptosis in osteoclasts. Recently it was shown anti-tumoral and anti-angiogenic effects of BPs, in vitro and in vivo. The present study aim to analyze and compare the anti-angiogenic pathway of the non-nitrogen-containing BP, clodronate (CLO), and the nitrogen-containing BP, zoledronic acid (ZOL), using estrogen receptor-positive breast cancer cell line, MCF-7, and endothelial cells. ZOL treatment reduced MCF-7 cell proliferation and induced cell cycle arrest in a dose-dependent manner, while CLO did not affect these cells. Treatment with ZOL or CLO also reduced RAEC cells proliferation, without changes in cell viability. HUVEC cells showed reduced viability after incubation with ZOL, but not with CLO. The conditioned medium by MCF-7 ZOL-treated cells reduced the formation of blood vessels in vitro and proliferation of HUVEC cells, together with lower endothelial cells proliferation derived from mouse aortic rings. Enzyme-linked immunoabsorbent assay showed reduction of ~25% in VEGFA levels, and increased amount of VEGFA165b isoform in the conditioned medium by ZOL-treated MCF-7 cells. The nitrogen-containing BP also modulated the expression of mRNAs for VEGF in MCF-7 treated cells. In all assays, CLO showed less anti-angiogenic properties when compared with ZOL. Data provided from this study amplify the knowledge of BPs actions, contributing for new approaches of bone metastases in malignant tumors / Mestrado / Estomatologia / Mestre em Estomatopatologia

Neuroblastoma, a neuroendocrine tumor, is the most common cancer in infancy. 75 % of those affected are under the age of 5. The disease is heterogeneous and survival rate is low.   Current treatment of neuroblastoma consists of surgery, radiation and chemotherapy, where the targets for the treatment are the malign cells. Due to the cancer cells instable genome there is a risk for resistance development. This negatively impacts the treatments goal of hindering tumor growth and spread.  Tumor growth is not only determined by malign cells but also the interactions of those tumor cells with tumor vessels and different types of cells in the tumor stroma.   The aim of this paper is to develop a relevant histological method to study the properties of tumor stroma in tumor sections retrieved from human NB tumor xenografts in mice treated with angiogenesis inhibitors SU11657 and Zoledronic acid. The study is a continuation of previous studies with the inhibitors which have shown good effect on tumor growth and angiogenesis on neuroblastoma.   In the short term treatment with SU11657 and Zoledron acid showed that tumor growth declined. In the longer treatment with SU11657 the growth didn’t decline with the same rate compared to the short term treatment. Angiogenesis on the other hand decreased in all the treatments independent of treatment duration. The histological staining with Sirius red revealed that treated tumors had an increased amount of stroma compared to the untreated tumors.   In conclusion the relative increase of tumor volume, decreased number of vessels and expansion of tumor stroma in the longer treatment with SU11657 indicated that tumors might survive the angiogenesis inhibitor treatment through expansion/activation of its stroma. The histological staining with Sirius red in saturated picric acid marked the collagen, i.e. stroma, well and enabled quantification of the stroma.

Large efforts have been made to target integrin αVβ3 of endothelial cells. We have successfully developed a new class of protein (Ref to as ProAgio) by rational protein design using a stable host protein, domain 1 of cell adhesion protein CD2. ProAgio is designed to target integrin αVβ3 at a novel site and induces angiogenic endothelial cell apoptosis by recruiting and activating caspase 8 to the cytoplasmic domain of the targeted integrins. Tests with tumor xenograft models show that ProAgio strongly inhibits tumor growth. Histology analyses indicate that tumor vessels are reduced, while the established vasculatures are not affected. Toxicity analyses demonstrate that ProAgio is not toxic to mouse. Our study develops an effective anti-angiogenesis agent and provides a new platform for development of therapeutics by targeting integrins. We have successfully developed an anti-angiogenesis protein targeting integrin αVβ3 at a novel site by rational protein design. The developed agent is not toxic to non-cancerous blood vessels and other tissue/organs, providing an excellent candidate for future potential clinical development. Our developed protein is one of the very few examples that do not act through targeting VEGF/VEGFR or any other RTK pathways. The βA groove is present in almost all other β integrins. This approach may be applicable to develop agents targeting the similar βA groove of other integrin pairs, which can address wide array of pathological conditions such as AMD, Rheumatoid Arthritis, Osteoporosis etc.

Le diabète est une maladie associée à une perturbation du métabolisme glucidique et représentent un problème de santé publique majeur. L'importante majorité des patients présente un diabète de type 2.Les complications les plus courantes sont vasculaires. Chez le diabétique, l'angiogenèse est défectueuse et paradoxale. La microangiopathie qui survient lors du diabète modifie le microenvironnement de la moelle osseuse et entraine egalement des problemes de cicatrisation alors qu'une angiogenèse exacerbé est responsable de pathologie telle que la rétinopathie ou la néphropathie diabétique.Les cellules souches mésenchymateuses (CSMs) sont connues pour leur potentiel de différenciation et de libération de facteurs paracrins, qui sont impliqués dans la régénération tissulaire. Les mécanismes qui associent le microenvironnement et les fonctions des CSMs dans un contexte diabétique restent actuellement méconnus. Le diabète peut modifier les caractéristiques des CSMs et le microenvironnement diabétique peut influencer la fonctionnalité de CSMs transplantées tout autant que leurs effets autocrins et/ou paracrins sur les cellules environnantes. L'étude du potentiel de CSMs diabétiques et du microenvironnement diabétique sur de CSMs pourrait alors avoir d'importantes implications cliniques. L'objectif primaire de cette étude est de caractériser l'impact du diabète sur la fonctionnalité et le potentiel angiogènique de CSMs à l'aide du modèle de rat dit Zucker Diabetic Fatty (ZDF) qui développe spontanément le diabète de type 2 et les complications vasculaire qui l'accompagne. / Diabetes is a disease associated with a bad functioning of glucose metabolism and represents a major health issue. The majority of diabetic are type 2 diabetic patients.Main complications in diabetes are vascular. Diabetic patients have paradoxical angiogenesis pathologies. Microangiopathy which occurs during diabetes modifies the microenvironment of bone marrow and lead to problem of wound healing whereas exacerbate angiogenesis is responsible for pathologies such as diabetic retinopathy or nephropathy.Mesenchymal stem cells (MSCs) are known for their differentiation potential and their release of bioactive mediators which are involved in tissular regeneration. The mechanism associated with microenvironment and functions of MSCs in a diabetic context remain elusive. Diabetes can change the characteristics of MSCs and diabetic microenvironment can lead to modifications in functionality of transplanted MSCs as well as modifying their autocrine/paracrine capacity on surrounding cells. The study of diabetic MSCs potential and the diabetic microenvironment on MSCs could have major clinical implications. The primary objective of this study was to characterize the impact of diabetes on functionality and angiogenic potential of MSCs with the help of a rat model, the Zucker Diabetic Fatty (ZDF) which develop spontaneously diabetes type 2 and the whole vascular complication associated with it.

Skeletal muscle is one of the largest tissues in humans and provides many pivotal functions to support life. Abnormality in skeletal muscle functions can lead to disease. For example, insulin resistance in skeletal muscle leads to type II diabetes. The underlying mechanisms that control energy balance in skeletal muscle remain largely elusive, especially at the genetic level. Here in the second chapter, I showed that MyoD mediated the transcriptional regulation of ACSL5, a mitochondrial protein, in C2C12 myoblasts via two E-box elements. A SNP rs2419621 (T) created a de novo E-box that together with the two pre-existing proximal E-boxes strongly enhances ACSL5 expression in both CV1 and C2C12 cells. In the third chapter, I identified a novel VGLL4-interacting protein IRF2BP2 and verified the interaction with co-immunoprecipitation and mammalian two-hybrid assays. Functionally, overexpression of IRF2BP2 and transcription factor TEAD1 activates mouse VEGF-A promoter in CV1 cells and enhances the biosynthesis of VEGF-A in C2C12 myoblasts. In vivo studies showed that ischemia induced the expression of IRF2BP2 by more than three fold, suggesting that IRF2BP2 could play a pivotal role during tissue ischemia. IRF2BP2 is a nuclear protein in both mouse cardiac myocytes and C2C12 myoblasts as demonstrated by immunohistochemistry and immunocytochemistry, respectively. Therefore, I sought to delineate the mechanism for the nuclear shuttling of IRF2BP2 in the fourth chapter. With various DNA alternations, I mapped the NLS to an evolutionarily conserved sequence 354ARKRKPSP361 in IRF2BP2. Deletion of the positively charged amino acids resulted in the abolishment of the NLS signal. Next, I showed that phosphorylation of serine 360 (S360) mediates the nuclear import of the protein. Whereas an alanine substitution (S360A) at the site resulted in perinuclear accumulation of the protein, an aspartic acid substitution (S360D) forced the nuclear accumulation. Nevertheless, the forced accumulation of the S360D mutant did not enhance the activation of VEGF-A promoter in CV1 cells as did the wild-type protein. My studies revealed two novel mechanisms by which skeletal muscle could harvest energy, thus providing new insight into the energy metabolism in skeletal muscle

Parmi les pathologies ischémiques, l'ischémie aiguë des membres inférieurs (IAMI) fait l'objet de nombreuses recherches ayant pour but une meilleure compréhension des mécanismes physiopathologiques et la mise au point de thérapies efficaces. Les cellules progénitrices endothéliales (PEC) participent à la régénération des vaisseaux lors d'événements ischémiques. En pathologie tumorale, la résistance aux traitements disponibles pousse la recherche à trouver de nouvelles cibles thérapeutiques. Depuis plusieurs années, notre équipe travaille sur la molécule CD146. Il a été démontré que la forme soluble de CD146 est une molécule angiogénique impliquée en physiologie et en pathologie. Notre travail a donc consisté à étudier le mécanisme d'action de cette molécule en pathologies. Les travaux de cette thèse comportent plusieurs axes :Un premier dans lequel l'étude des modulations des effets de CD146 soluble sur les PEC a permis de mettre en évidence son récepteur, l'angiomotine. La seconde partie du travail a porté sur l'étude des effets d'un prétraitement par CD146 soluble de PEC sur un modèle d'IAMI chez la souris. In vitro et in vivo, CD146 soluble augmente les propriétés angiogéniques et la viabilité des PEC.Enfin, la troisième partie des travaux réalisés durant ma thèse a porté sur le rôle de sCD146 en pathologie cancéreuse. Nous avons développé des modèles de xénogreffes decellules cancéreuses nous permettant d'examiner les effets de CD146 soluble sur la croissance tumorale par l'injection de la molecule. Les résultats obtenus montrent que CD146 soluble augmente la croissance et la vascularisation tumorale. / Diseases with angiogenic component such as ischaemic pathologies and cancer have a high incidence. Among ischaemic pathologies, the acute ischaemia of the lower limbs made the object of many research having for goal a better comprehension of the physiopathological mechanisms and the development of effective therapies. The endothelial progenitor cells (EPC) take part in the regeneration of the vessels during ischaemia. In tumoral pathology, resistance to available treatments pushes research to find new therapeutic targets. For several years, our team has worked on CD146 molecule. It was shown that the soluble form of CD146 is an angiogenic factor involved in physiology and pathology. Our work thus consisted in studying the mechanism of action of this molecule in pathologies. Work of this thesis comprises several axes: A first in which the study of the modulations of the effects of soluble CD146 on EPC made it possible to highlight its receptor, angiomotin protein. The second part of the work concerned the study of the effects of a pretreatment by soluble CD146 on EPC on a model of IAMI in mouse. In vitro and in vivo, soluble CD146 increases the angiogenic properties and the viability of the EPC. Lastly, the third part of the work completed during my thesis concerned the role of sCD146 in cancerous pathology. We developed xenograft models of cancer cells allowing us to examine the effects of soluble CD146 on the tumor growth by the injection of this molecule. The results obtained show that soluble CD146 increases tumor growth and vascularization.

Clinical manipulation of angiogenesis (the formation of new blood vessels from pre-existing vasculature) is of interest to treat diseases such as cancer and ischemic tissue where it is not properly regulated. Several treatments targeting vascular endothelial growth factor (VEGF) and its receptors - which are abundant at sites of angiogenesis - are currently in use to treat various types of cancer, however they have severe vascular side effects. Conversely, VEGF has been used clinically to promote angiogenesis to treat ischemic tissue. However, despite encouraging data from pre-clinical models, trials in humans have been disappointing. For further therapies to be developed, more information on how VEGF interacts with its receptors is required. Heparan sulfate (HS) is a ubiquitous glycosaminoglycan involved in a number of physiological processes including angiogenesis. HS facilitates the interaction of VEGF with its receptors, which is crucial for angiogenesis. Modification of this interaction via synthetic mimetics of HS may allow clinical intervention of angiogenesis. The current investigation aims first, to clarify the requirement for the interaction between VEGF and HS in angiogenesis; second to characterise the structure of HS that binds to VEGF so that mimetics can be developed; and third, to determine the effect of HS mimetics on angiogenesis in vivo. To determine the requirement for VEGF/HS interaction in angiogenesis, several mutants of VEGF165 that had lower affinities for HS were assayed for their ability to induce ectopic angiogenesis in the subintestinal baskets of zebrafish embryos. Wild type VEGF165 induced a 200-250% increase in ectopic vessels, which was matched only by a control mutant. Other mutants did not induce ectopic vessels, suggesting that this interaction is required for angiogenesis. To characterise the structure of HS that binds to VEGF, various HS mimetics were assayed against heparin in a VEGF competition assay using Biacore. Of these, the strongest inhibition (IC¬50 =~16nM) was with 2O10, an oligosaccharide that consisted of two highly sulfated octasaccharide domains (NS domains) that flanked an unsulfated dodecasaccharide region. To determine the type of sulfation required for this interaction, HS fragments were assayed for interaction with VEGF165 using the filter binding assay, and analysed by HPLC which indicated 6-O sulfation may be preferential for VEGF binding to HS.To investigate the ability of HS to affect angiogenesis, the effects of HS mimetics on zebrafish embryo subintestinal baskets were measured. The most interesting of these was with 2O10, which had a biphasic response whereby low doses (3ng) increased basket vasculature by 30% and high doses (30ng) decreased the endogenous vessels by 20%. As 2O10 had a high affinity for VEGF, its effects on the vasculature may be due to interaction with endogenous VEGF, which would indicate that HS mimetics can be used to control angiogenesis by modification of growth factor signalling. The investigation concludes that the interaction between VEGF and HS is critical for angiogenesis, and that this can be modulated by the application of HS mimetics that bind strongly to VEGF.

O objetivo deste trabalho foi avaliar a presença da proteínas - fator de crescimento endotelial vascular (VEGF) e seus receptores R1 (Flt-1) e R2 (Flk-1) em placentas a termo de gestantes com hipergliccemia leve, comparando estes resultados com o observado em placentas de gestantes normoglicêmicas e diabéticas gestacional e clínica, através de reações de imunohistoquímica e Western Blotting. Resultados: em geral, placentas de gestantes normoglicêmicas apresentam reatividade bastante expressiva aos anticorpos contra VEGF, VEGF-R1 e -R2 nas células vasculares e trofoblásticas. Reações destacadamente intensas foram observadas no endotélio capilar, células mesenquimais, sinciciotrofoblasto e células citotrofoblásticas extravilosas para VEGF e VEGF-R2. Estes resultados sugerem que o balance VEGF/VEGFR está alterado nas placentas de gestantes com hiperglicemia leve, o que pode ser um aspecto crucial para explicar a hipercapilarização induzida nos vilos terminais neste distúrbio glicêmicos. / O objetivo deste trabalho foi avaliar a presença da proteínas - fator de crescimento endotelial vascular (VEGF) e seus receptores R1 (Flt-1) e R2 (Flk-1) em placentas a termo de gestantes com hipergliccemia leve, comparando estes resultados com o observado em placentas de gestantes normoglicêmicas e diabéticas gestacional e clínica, através de reações de imunohistoquímica e Western Blotting. Resultados: em geral, placentas de gestantes normoglicêmicas apresentam reatividade bastante expressiva aos anticorpos contra VEGF, VEGF-R1 e -R2 nas células vasculares e trofoblásticas. Reações destacadamente intensas foram observadas no endotélio capilar, células mesenquimais, sinciciotrofoblasto e células citotrofoblásticas extravilosas para VEGF e VEGF-R2. Estes resultados sugerem que o balance VEGF/VEGFR está alterado nas placentas de gestantes com hiperglicemia leve, o que pode ser um aspecto crucial para explicar a hipercapilarização induzida nos vilos terminais neste distúrbio glicêmicos.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22040号 / 医博第4525号 / 新制||医||1038(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 高橋 淳, 教授 伊佐 正, 教授 渡邉 大 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM