Common measures of egg quality have been survival to specific developmental stages, higher hatching rate of fertilized eggs and final production of fry. Determinants of egg quality are variable among and between teleost species and no common unified criteria have been established. Maternally inherited genes influence egg quality and early embryo development is partially programmed by the messenger ribonucleic acid (mRNA). Among the genes, the insulin family is important for growth functions and the presence of their transcripts in the ovary, oocytes and embryos implies their involvement during the reproductive process and their relevance to egg quality. The insulin-like growth factor (IGF) system has three components, the ligands IGF-I and II, the IGFBPs (insulin-like growth factor binding proteins) and the IGF receptors that mediate biological activity of the ligands. Vitellogenin (Vtg) is the major source of nutrients for the developing embryo and elevated levels in female fish plasma signals gonadal development preceding spawning. In oviparous fish where the developing embryo is dependent on the stored food in the yolk, vitellogenin levels in the egg could indicate its capability to support embryonic growth. This study aimed to develop molecular tools, specifically probes for IGF-I, IGF-II and IGF-IR, for the evaluation of fish egg quality. These probes would be used to determine expression levels of IGF-I, IGF-II and IGF-IR during egg development to assess their potential as molecular indicators for egg quality. In addition, this study also aimed to establish an enzyme-linked immunoassay (ELISA) for quantifying Vtg in fish eggs and determine if differences in Vtg levels could be linked to fertilization and hatching success. Through reverse-transcription polymerase chain reaction (RT-PCR) putative complementary deoxyribonucleic acid (cDNA) fragments of IGF-I, IGF-II and IGF-IR were cloned and sequenced from mullet (Mugil cephalus) and grouper (Epinephelus coioides). The relative expression ratio of the three genes in the eggs of mullet and grouper were assayed by quantitative PCR (QPCR) and calculated using the Pfaffl method (Pfaffl, 2001). Levels of vitellogenin in different batches of mullet eggs were quantified by ELISA. Spawned eggs of grouper were grouped into low (<60%) or high (>60%) fertilization rate (FR) and the fertilized eggs that were incubated until hatching were grouped into medium (>90%) or high (>90%) hatching rate (HR). Samples were categorized into sinking eggs, late embryo and hatched larvae. Relative expression ratio of IGF-II was significantly high (P<0.01) compared to IGF-I and IGF-IR in all samples examined. All three genes were strongly expressed in sinking eggs compared to either late embryo or hatched larvae. However, there was no significant interaction effect between the genes and the samples analyzed. Mullet samples all came from a high FR and high HR group and were categorized into sinking, multicell stage, blastula, gastrula, late embryo and hatched larvae. There was a significant interaction effect (P<0.01) between gene and stage, showing that genes are differentially expressed during embryonic development. IGF-II was strongly expressed relative to the other genes in all stages examined and was highest during the gastrula stage. Vtg levels were examined in mullet oocytes and egg samples that were grouped into 4; oocytes from females that subsequently spawned, had fertilized eggs which hatched (Group A); oocytes from females that did not spawn, therefore no fertilization and no hatching (Group B); eggs that were stripped, artificially fertilized but no hatching (Group C); and eggs that were spawned, assumed to be fertilized but did not hatch (Group D). Group A showed a trend of higher Vtg levels than the other three but this result was not statistically significant.

O tecido adiposo e ósseo tem uma íntima relação, desde a origem comum nas células tronco estromais derivadas da medula óssea. Sabe-se que o peso corporal tem estreita correlação com a massa óssea em seres humanos. Porém, ainda não é claro qual componente do peso corporal tem maior influência sobre o ganho de massa óssea e sobre a adiposidade na medula óssea, visto que tanto indivíduos com baixo peso, quanto os obesos, apresentam altas taxas de fraturas. O objetivo deste trabalho foi comparar crianças e adolescentes obesos e eutróficos em relação a composição óssea, adiposidade da medula óssea em coluna lombar (L3), expressão do Receptor tipo 1 de IGF (IGF1R) e concentrações séricas de IGF-I e buscar correlação entre estas variáveis. Para tanto foram avaliados crianças e adolescentes de 10 a 17 anos, divididos em grupo controle e grupo obeso. Esses grupos foram submetidos a avaliação antropométrica, densitometria óssea de coluna lombar e corpo total e ressonância magnética de coluna lombar e abdome total, além de dosagens séricas de parâmetros bioquímicos e hormonais. Os pacientes do grupo obeso apresentaram associação positiva da densidade mineral óssea tanto com massa gorda quanto com massa magra, enquanto que o grupo controle apresentou associação positiva da densidade mineral óssea apenas com a massa gorda. Não houve diferença entre os grupos quanto a adiposidade da medula óssea, nem quanto aos valores de IGF-I, IGFBP3 e expressão do gene do IGF1R. / The adipose and bone tissue has an intimate relationship, from the common origin in stromal stem cells derived from the bone marrow. It is known that body weight has a close correlation with bone mass in humans. However, it is still unclear which component of body weight has a greater influence on bone mass gain and adiposity in the bone marrow, since both individuals with low weight and obese have high fracture rates. The objective of this study was to compare obese and eutrophic children and adolescents in relation to bone composition, bone marrow adiposity in the lumbar spine (L3), expression of IGF type 1 receptor (IGF1R) and serum concentrations of IGF-I and to seek correlation between these variables. For this, children and adolescents between 10 and 17 years old were divided into control and obese groups. These groups were submitted to anthropometric evaluation, bone densitometry of the lumbar spine and total body and lumbar spine and total abdominal magnetic resonance, in addition to serum levels of biochemical and hormonal parameters. The patients in the obese group had a positive association of bone mineral density with both fat mass and lean mass while the control group showed a positive association of bone mineral density with fat mass only. There was no difference between the groups regarding bone marrow adiposity, nor regarding IGF-I, IGFBP3 and IGF1R gene expression.

IGF-I induz proliferação e diferenciação celular. Neste estudo visamos sua participação na imunidade específica na leishmaniose. Estudamos efeito de citocinas Th1 (IFN-g) e Th2 (IL- 4 mais IL-13) na modulação da produção de IGF-I e seu reflexo no parasitismo de macrófago (MØ) de camundongos BALB/c e C57BL/6, infectados por amastigotas e promastigotas de Leishmania (L.) major. Iniciamos avaliando os efeitos de IGF-I no modelo, onde no desenvolvimento da lesão IGF-I induziu aumento maior de lesão em camundongos BALB/c. Nos camundongos C57BL/6 o efeito foi marcante com evolução progressiva da lesão enquanto que sem o fator, a lesão se controlava. Como a produção de IGF-I por MØ é modulada por citocinas, onde IFN-g diminui e IL-4 e IL-13 aumentam a sua expressão, fomos estudar seus efeitos no modelo proposto onde MØ foram infectados com amastigotas ou promastigotas de L.(L.) major (parasitos/célula = 2:1) e incubados com IFN-g (200U/mL) ou IL-4 (2ng/mL) mais IL-13 (5ng/mL). IFN-g induziu aumento significante na produção de NO nos dois modelos estudados. O efeito de citocinas Th1 e Th2 sobre o parasitismo em MØ foi distinto em BALB/c e C57BL/6. Quando infectados por amastigotas, o parasitismo aumentou sob estímulo com IL-4 mais IL-13 e diminuiu com IFN-g como era esperado. No entanto, quando infectado por promastigota, o parasitismo aumentou com IL-4 mais IL-13 somente em células BALB/c. Por outro lado, o parasitismo diminuiu com IFN- g somente em células C57BL/6. Mesmo na ausência do estímulo por citocinas, observou-se aumento na expressão de RNA de IGF-I nos MØ de animais BALB/c infectados por formas amastigotas ou promastigotas e de C57BL/6 infectados por promastigotas. Um resultado destoante foi à diminuição da expressão de IGF-I em MØ de C57BL/6 infectados por amastigotas. Em células BALB/c, essa expressão aumentada de IGF-I ocorrida em função da infecção, somente se alterou com IL-4 mais IL-13 quando infectadas por amastigotas. Em células C57BL/6, a expressão aumentada de IGF-I com a infecção por prormastigota, sofreu diminuição sob estímulo com IFN-g, não se alterando com citocinas Th2. A expressão diminuída de IGF-I com a infecção por amastigotas diminuiu ainda mais com IFN-g e aumentou com citocinas TH2. Os dados sugerem a possibilidade de modulação do efeito de citocinas pela expressão diferenciada de IGF-I em macrófagos infectados por L. (L.) major / Insulin-like growth factor (IGF)-I induces cell proliferation and differentiation. In this study, we focused on its participation on specific immunity. We studied the effect of Th1 (IFN-g) and Th2 (IL-4 plus IL-13) cytokines on the modulation of the production of IGF-I and its influence on BALB/c and C57BL/6 macrophage (MØ) parasitism using Leishmania (L.) major amastigote and promastigote. IGF-I expression was increased in infected macrophages. Initially, we substantiated the effect of IGF-I on in vitro growth of Leishmania (L.) major promastigote, on cutaneous lesion development in vivo and diminished production of nitric oxide in MØ. On development of the lesion, IGF-I induced greater increase of the lesion in BALB/c mice. In C5BL/6 mice, its effect was more pronounced with progression of the lesion whilst without the factor it was controlled. Since IGF-I production by MØ is modulated by cytokines where IFN-g decreases and IL-4 plus IL-13 increase its expression, we studied their effects in BALB/c and C57BL/6 MØ. Macrophages were infected with L.(L.) major amastigotes or promastigotes (parasites/cell = 2:1) and incubated with IFN-g (200U/mL) or IL-4 (2ng/mL) plus IL-13 (5ng/mL). IFN-g induced a significant increase of NO production by MØ from both models. The effect of Th1 and Th2 cytokines on parasitism was distinct when MØ infected with amastigotes, the parasitism increased upon IL-4 plus IL-13 stimuli and diminished with IFN-g as expected. However, when infected with promastigotes, the parasitism increased with IL-4 plus IL-13 only in BALB/c cells. Conversely, the parasitism diminished with IFN-g only in C57BL/6. Even in the absence of cytokine stimuli, an increase of the expression of IGF-I mRNA was observed in L (L.) major amastigote- and promastigote- infected BALB/c MØ and in promastigoteinfected C57BL/6 MØ. A dissonant result was a diminished IGF-I expression in amastigoteinfected C57BL/6 MØ. In BALB/c this increased IGF-I expression with infection only altered with IL-4 plus IL-13 in amastigote-infected cells. In C57BL/6 cells, the increased IGF-I expression upon prormastigote infection diminished under IFN-g stimulus that was not altered with Th2 cytokines. A decreased IGF-I expression with amastigote-infection diminished even more with IFN-g and increased with Th2 cytokines. These data suggest a possibility of modulation of the effect of cytokines on distinct expression of IGF-I in L. (L.) major-infected MØ

Made available in DSpace on 2014-08-20T13:32:53Z (GMT). No. of bitstreams: 1 dissertacao_augusto_schneider.pdf: 2025669 bytes, checksum: bde04bd1476df3f35564859c10477bca (MD5) Previous issue date: 2008-11-28 / In non-ovarian tissues GHR expression is regulated by insulin concentrations and is correlated to IGF-I expression. In the ovary IGF-I expression is also related to insulin levels, however no correlation with GHR was demonstrated until now. The aim of this study was to investigate the effect of fasting or insulin administration on blood concentrations of glucose, nonsterified fatty acids, insulin, insulin like growth factor I (IGF-I) and estradiol and on follicular expression of growth hormone receptor (GHR) and IGF-I mRNA in ewes. Fifteen ewes received an intravaginal progesterone releasing device that was removed 6 days later (Day 0). In Day -2 the ewes were divided in: control group, which received maintenance diet, insulin group, which received insulin injections every 12 hours from Day -2 to 2 and fasting group, which was submitted to fasting from Day -2 to 2. The results of the current study revealed that insulin administration or fasting during the development of a follicular wave did not affect (P=0.22) follicular diameter, although insulin injection increased (P=0.02) estradiol production. There was no difference among groups for GHR or IGF-I mRNA expression in granulosa (P=0.62, 0.43) or theca cells (P=0.92, 0.43). For fasting group there was a positive correlation between glucose and estradiol (r=0.97, P=0.006) and granulosa cell IGF mRNA (r=0.96, P=0.03). For insulin group estradiol was positive correlated to follicular diameter (r=0.93, P=0.02) and granulosa cell GHR (r=0.87, P=0.05) and IGF-I mRNA (r=0.79, P=0.10). In conclusion, exogenous insulin or fasting did not affect follicular diameter and expression of GHR and IGF-I mRNA in the pre-ovulatory follicle, although exogenous insulin increased estradiol production. / Em tecidos não ovarianos a expressão do receptor do hormônio do crescimento (GHR) é regulada pelas concentrações de insulina e está correlacionada a expressão de fator de crescimento semelhante a insulina tipo I (IGF-I). No ovário a expressão de IGF-I também está relacionada aos níveis de insulina, porém ainda não foi demonstrada sua correlação com a expressão de GHR. O objetivo deste estudo foi investigar o efeito do jejum ou administração de insulina nos níveis de glucose, ácidos graxos não esterificados, uréia, IGF-I, insulina e estradiol e na expressão folicular de RNAm para GHR e IGF-I em ovelhas. Quinze ovelhas receberam um dispositivo intravaginal liberador de progesterona, que foi removido após seis dias (Dia 0). No Dia -2 as ovelhas foram dividas em: grupo controle, que recebeu uma dieta de manutenção; grupo insulina, que recebeu injeções de insulina a cada 12 horas do Dia -2 ao Dia 2 e grupo jejum, que foi submetido à dieta hídrica do Dia -2 ao Dia 2. Os resultados revelaram que a administração de insulina ou o jejum durante o desenvolvimento de uma onda folicular não influenciaram (P=0,22) o diâmetro folicular, no entanto a administração de insulina aumentou (P=0,02) a produção de estradiol. Não houve diferença entre os grupos para a expressão de RNAm para GHR e IGF-I nas células da granulosa(P=0,62; 0,43) ou teca (P=0,92; 0.43). Para o grupo jejum houve uma correlção positiva entre glucose e estradiol (r=0,97, P=0,006) e RNAm IGF-I nas células da granulosa (r=0,96, P=0,03). Para o grupo insulina o estradiol foi positivamente correlacionado ao diâmetro folicular (r=0,93, P=0,02) e expressão de RNAm para GHR (r=0,87, P=0,05) e IGF-I (r=0,79, P=0,10)nas células da granulosa. Em conclusão, a insulina exógena ou o jejum não afetaram o diâmetro folicular e a expressão de mRNA para GHR e IGF-I no folículo pré-ovulatório, apesar da insulina exógena ter aumentado a produção de estradiol.

A insuficiência cardíaca (IC) é a via final comum da maioria das cardiomiopatias e outras doenças do aparelho circulatório. Considerando a prevalência crescente e a morbimortalidade associada representa um importante problema de saúde pública. Em quadros mais avançados, além do comprometimento funcional, portadores de IC apresentam perda de massa muscular excessiva que pode culminar em caquexia cardíaca; condição que contribui para o mau prognóstico e a mortalidade aumentadas. A massa muscular é regulada pelo balanço entre estímulos anabólicos e catabólicos. A quinase Akt vêm sendo considerando uma importante quinase na regulação do crescimento muscular por controlar o anabolismo proteico. Dessa forma, ativadores da Akt (IGF-I e insulina), bem como proteínas alvo da sinalização da Akt (mTOR e GSK3) são importantes mediadores na manutenção da massa muscular e podem ser regulados por estímulos metabólicos, nutricionais e mecânicos. Assim, o objetivo desse estudo foi avaliar a contribuição da via de sinalização IGF-I/Akt/mTOR na atrofia muscular desencadeada pela IC tanto em humanos quanto em modelo experimental, bem como o efeito do treinamento físico aeróbico (TFA). Nossos resultados demonstraram que em biópsias do vasto lateral de pacientes portadores de IC classe II houve redução na expressão de mRNA de todas as isoformas de IGF-I e na expressão das proteínas IGFBP-3, Akt1, GSK3, pGSK3Ser9, mTOR e tendência a redução na pmTORSer2448 (p=0,08) e aumento na pAMPKThr172. Esses resultados foram acompanhados pela redução no VO2 pico desses pacientes. O TFA de 12 semanas levou a um aumento não significativo na expressão de mRNA da isoforma IGF-I Ea (p=0,07) e IGF-I PAM (p=0,06). Também observou-se aumento na pAMPKThr172 e tendência a aumento na Akt1 (p=0,07) e mTOR (p=0,06). Em modelo experimental de IC, no músculo sóleo observou-se redução na expressão das proteínas IGF-I, PI3K, pAktSer473,pGSK3Ser9 e aumento da pAMPKThr172. O TFA de 8 semanas promoveu aumento na expressão do mRNA das isoformas de IGF-I Ea, Eb e IGF-I PAM, bem como na expressão das proteínas IGFI, PI3K, pAktSer473, pmTORSer2448 e redução na pAMPKThr172. O conjunto de alterações promovido pelo TFA foi associado à maior tolerância ao esforço físico e ganho no desempenho motor em Rota Rod, além de prevenir a atrofia muscular. Quando esses animais foram tratados com rapamicina, um inibidor farmacológico da mTOR, o efeito do TFA na prevenção da atrofia muscular foi abolido. Juntos, esses resultados apoiam a hipótese de que a via de sinalização IGF-I/Akt/mTOR está envolvida no reestabelecimento muscular na IC induzida pelo TFA / Heart failure (HF) is a common ultimate consequence for the most cardiomyopathies and other diseases from circulatory system. Due its growing prevalence and morbimortality is an important public-health problem. In more advanced cases, besides functional impairment, HF patients present an excessive skeletal muscle loss which can lead to cardiac cachexia; a condition associated to a poor prognostic and increased mortality. Skeletal muscle mass is regulated by the balance between anabolic and catabolic stimuli. Akt kinase, although involved with the protein synthesis pathway, is able to regulates the skeletal muscle growth via anabolism and catabolism control. An importante role in the skeletal muscle growing has been attributed to the Akt kinase due its ability to control protein synthesis. Thus, activators (IGF-I and insulin) as well as target proteins in the Akt signaling pathway (mTOR and GSK3) are important mediators in the skeletal muscle mass homeostasis being regulated by anabolic, nutritional and mechanic stimuli. Therefore, the aim of the presente study was to evaluate the contribution of IGF-I/Akt/mTOR contribution to the muscle atrophy induced by HF in humans and mice. Additionally, the effect of aerobic exercise training were evaluated. Our data demonstrated that in biopsies obtained in the vastus lateral from class II IC patients occurred a reduction in the expression of all the forms of IGF investigated and in the expression of proteins such as IGFBP-3, Akt1, GSK3, pGSK3Ser9, mTOR, and tendency to reduce pmTORSer2448 (p=0.08) and to increase pAMPKThr172. These results were accompained by the reduction of peak VO2 in these patients. Twelve weeks of aerobic exercise training promoted a non-significant increase in the expression of the IGF-I Ea (p=0.07) and IGF-I PAM (p=0.06) mRNA expression. Moreover, it was observed an increase to pAMPKThr172, and tendency of increase for Akt1 (p=0.07) and mTOR (p=0.06) mRNA expression. In a experimental model of IC, it was observed a reduction in the expression of IGF I, PI3K, pAktSer473, pGSK3Ser9 and increase of pAMPKThr172. Eight weeks of aerobic exercise training increased the mRNA of IGF-I Ea, Eb and IGF-I PAM isoforms, as well as in the expression of IGF-I, PI3K, pAktSer473, pmTORSer2448 and pAMPKThr172 reduction. The changes induced by aerobic exercise training were associated with a higher tolerance to exercise and motor performance in rota rod besides prevent muscular atrophy. When treated with a inhibitor of mTOR, rapamycin, the adaptations induced by aerobic exercise training were blunted, supporting the hypothesis that the IGF-I/Akt/mTOR signaling pathway is involved in the recovering of muscle mass induced by physical training

Die im Rahmen dieser Arbeit durchgeführten Untersuchungen zum Einfluss der Wachstumsfaktoren IGF-I, EGF und dem Zytokin IL-6 auf den Androgenrezeptor-Signalweg zeigten in verschiedenen Prostatakarzinom-zelllinien schon nach zwei Stunden eine deutliche Degradation des Androgenrezeptor-Proteins. Die ausschließlich auf Protein-Ebene stattfindende, Wachstumsfaktor-induzierte negative Regulation des Androgenrezeptors konnte durch einen schnellen Androgeneffekt wieder aufgehoben werden. Mittels Luziferase-Reportergen-Assays wurde kein Einfluss der Wachstums-faktorwirkung auf die transkriptionelle Aktivität des Androgenrezeptors nachgewiesen. Darüber hinaus konnte eine signifikant reprimierende Wirkung durch IGF-I und EGF in Kombination mit geringen Mengen DHT beobachtet werden. Weitere Resultate dieser Arbeit deuten auf einen, durch den PI3-Kinase-Signalweg vermittelten, proteasomalen Abbauprozess des Rezeptors hin. Da die Suppression der downstream gelegenen Proteinkinase Akt keine Veränderung hinsichtlich der Degradation aufwies, konzentrierte sich die weiterführende Arbeit auf eine mögliche direkte Regulation des Androgen-rezeptors durch die PI3-Kinase. Unter Verwendung von rekombinanten GST-Fusionsproteinen konnte in Interaktionsstudien unter in vitro Bedingungen eine Phosphotyrosin-unabhängige Bindung zwischen der C-SH2-Domäne der p85-Untereinheit der PI3-Kinase und dem N- und C-Terminus des Androgenrezeptors nachgewiesen werden. Durch die nähere Charakterisierung dieser Bindungsbereiche mit Hilfe von Peptidarrays und anschließenden Alanin-Substitutionen war es möglich, für den N-Terminus 18, für den C-Terminus des Androgenrezeptors 6 und für die p85-C-SH2-Domäne der PI3-Kinase 11 Aminosäuren zu identifizieren. Die durch gezielte Punktmutagenese an diesen Aminosäurepositionen hergestellten Androgenrezeptor-Einzel- und -Mehrfachmutanten wiesen in Bindungsstudien dennoch Interaktion zur PI3-Kinase auf. Eine von Anderson und Kollegen postulierte Phosphotyrosin-unabhängige Bindung der SH2-Domänen der p85-Untereinheit der PI3-Kinase durch sogenannte „basic-X-basic“-Motive wurde ebenfalls in Interaktionstests zwischen der PI3-Kinase und dem Androgenrezeptor überprüft. Aufgrund der Tatsache, dass einige der identifizierten Aminosäuren auf dem Androgenrezeptor Teil eines „basic-X-basic“-Bindungsmotives sind, wurden Kombinationsmutanten generiert, die sowohl im N-Terminus als auch im C﷓Terminus des Androgenrezeptors ein bzw. zwei zerstörte „basic-X-basic“-Motive enthielten. Untersuchungen zum Bindungsverhalten dieser Mutanten zeigten zwar weiterhin Interaktion zur p85-C-SH2-Domäne der PI3-Kinase, jedoch der durch Western-blot-Analyse überprüfte IGF-I-induzierte Degradationseffekt des Androgenrezeptor-Proteins konnte mit zwei der verwendeten Androgenrezeptor-Kombinationsmutanten nicht mehr beobachtet werden.

Growth plate injuries account for 15-30% of long bone fractures in children. About 10% of these result in significant growth disturbances due to formation of a boney bar. If not treated correctly, this can lead to life-lasting consequences of limb length inequalities and angular deformities. Current treatments for growth plate injuries include removal of boney bar and insertion of fat, silicone, bone cement, etc.. This treatment y is inadequate, leaving almost half of these patients with continued deformities. This dissertation reports characterization of a DNA–containing porous poly(lactic-co-glycolic acid) (PLGA) scaffold system, chondrogenesis using insulin-like growth factor I (IGF-I) plasmid-releasing scaffolds in vitro, and in vivo testing of IGF-I plasmid-releasing scaffolds to regenerate growth plate . Controlled release of naked and DNA complexed with polyethylenimine (PEI) was achieved from porous PLGA scaffolds. PEI affected release of complexes from PLGA scaffolds, as PEI:DNA complexes were released at a lower rate compared to naked DNA encapsulated in low molecular weight (LMW) and high molecular weight PLGA scaffolds, as well as hydrophilic and hydrophobic PLGA scaffolds. Hydrophilicity and molecular weight of PLGA affected the release profiles of both naked DNA and PEI:DNA complexes from the scaffolds, as evidenced by later peak DNA and PEI:DNA release with increasing hydrophilicity and molecular weight. LMW hydrophilic PLGA scaffolds supported growth and chondrogenic differentiation of mesenchymal multipotent D1 cells, chondrocytes, and bone marrow cells (BMCs) in vitro. Culturing BMCs on IGF-I plasmid-encapsulated scaffolds resulted in elevated expression of IGF-I compared to blank scaffolds. Removal of boney bar and implantation of IGF-I plasmid-releasing LMW PLGA scaffolds in a rabbit model of growth plate injury resulted in some improvement of leg angular deformity compared to no scaffold implantation. Histological analysis of the newly developed cartilage showed growth plate-like columnar arrangement of chondrocytes in a defect that received IGF-I plasmid encapsulated scaffold, although the level of organization of newly formed cartilage was inferior to that of native growth plate. This appears to be the first report of the regeneration of growth plate-like structure without the use of stem cells in an animal model of physeal injury.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Made available in DSpace on 2014-06-11T19:30:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-10-04Bitstream added on 2014-06-13T20:00:58Z : No. of bitstreams: 1 leme_jaca_dr_rcla.pdf: 1133844 bytes, checksum: 96fcdb51142a4b6ddf31ece437a39c75 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Para investigar os efeitos do treinamento físico moderado nas concentrações de grelina, leptina, GH e IGF-I em ratos diabéticos, ratos wistar foram distribuídos nos seguintes grupos: controle sedentário, controle treinado, diabético sedentário e diabético treinado. A indução do diabetes foi feita por aloxana (32 mg/kg). Após determinação da carga equivalente à máxima fase estável do lactato (MFEL), o protocolo de treinamento físico de natação consistiu de 1 hora/dia, 5 dias/semana durante 8 semanas suportando uma carga equivalente à 90% da MFEL. Durante o período experimental foram registradas semanalmente a massa corporal e ingestão hídrica e alimentar. Ao final do período experimental, foi feita a eutanásia dos animais e o sangue foi coletado para determinação das concentrações de glicose, insulina, albumina, triglicerídeos, GH, IGF-I, grelina acilada e leptina. A massa do estômago foi registrada e foram coletadas amostras dos seguintes tecidos: fígado para determinação de IGF-I, proteína, DNA e triglicerídeos; pâncreas para determinação de IGF-I, proteína e DNA; hipófise para determinação de GH, estômago para determinação de grelina, proteína e DNA e tecido adiposo epididimal para determinação de leptina, proteína e DNA. O diabetes experimental reduziu a massa e comprimento corporal além das concentrações no sangue de insulina, leptina, GH e IGF-I, mas aumentou as ingestões hídrica e alimentar, e as concentrações de triglicerídeos, glicose e grelina acilada sanguíneas. Nos animais diabéticos, o exercício físico promoveu redução da ingestão alimentar, diminuiu as concentrações sanguíneas de glicose e triglicerídeos e recuperou os valores de IGF-I. No fígado, o diabetes reduziu os valores de IGF-I, proteína e razão proteína/DNA, aumentou os valores de triglicerídeos que foram recuperados pelo treinamento físico... / In order to investigate the effects of moderate physical training in the concentrations of ghrelin, leptin, GH and IGF-I in diabetic rats, Wistar rats were distributed in four groups: sedentary control, trained control, sedentary diabetic and trained diabetic. Experimental diabetes was induced by alloxan (32 mg / bw). After workload determination equivalent to maximal lactate steady-state (MLSS), the exercise training protocol consisted in swimming 1 h / day, 5 days / week during 8 weeks supporting a load equivalent to 90% of the MLSS. During the experimental period, weekly body mass, and water and food intake were recorded. At the end of the experiment, euthanasia was performed and the blood was collected to determine the level of glucose, insulin, albumin, triglycerides, GH, IGF-I, acylated ghrelin and leptin. The weight of the stomach was recorded and samples of the following tissues were collected: liver to determine IGF-I, protein, DNA and triglycerides; pancreas to determine IGF-I, protein and DNA; pituitary to determine GH; stomach to determine of ghrelin, protein and DNA and epididymal adipose tissue to determine leptin, protein and DNA. In addition to body weight and length, experimental diabetes also reduced blood concentrations of insulin, leptin, GH and IGF-I. Nevertheless, it increased water and food intake, and also the concentrations of triglycerides, blood glucose and acylated ghrelin. In diabetic rats, exercise caused a reduction of food intake, blood concentrations of glucose and triglycerides, and recovery blood IGF-I. In the liver, diabetes reduced the IGF-I, protein and protein/DNA ratio values, and increased triglycerides level, which was recovered by physical training in these parameters. Diabetes also reduced pituitary GH concentrations, which did not alter by physical training. On the other hand, training increased IGF-I and protein / DNA ratio values... (Complete abstract click electronic access below)

Background: The aim of this thesis was to study the relationship of pre-diagnostic circulating concentrations of sex steroid hormones (androgens, estradiol, 17-hydroxyprogesterone, and progesterone), growth factors (insulin-like growth factor-I (IGF-I), placental growth hormone (GH)), sex hormone binding globulin (SHBG), and anti-Müllerian hormone (AMH) with risk of epithelial ovarian cancer (EOC) overall, and by tumor invasiveness and histology. A longitudinal study was used to assess patterns of hormonal changes during a single pregnancy, and in two consecutive pregnancies. Materials &amp; Methods: A case-control study was nested within the Finnish Maternity Cohort and the Northern Sweden Maternity Cohort. A total of 1 052 EOC cases were identified through linkages with the cancer registries in both countries. For each case, 2-3 controls were selected. Cases and controls were matched on cohort, age and date at blood draw, as well as for parity at blood draw and at diagnosis (n=2 695). Odds ratios (OR) and corresponding 95% confidence intervals [CI] were estimated using conditional logistic regression. The longitudinal study was based on 71 pregnant Finnish women, who donated blood samples in each trimester of pregnancy. Results: Higher androgen concentrations were associated with an increased risk of overall EOC (e.g., testosterone ORT3 vs. T1: 1.56 [1.30-1.87], ptrend&lt;0.0001), while the risk of endometrioid tumors increased with higher estradiol concentrations (ORT3 vs. T1: 2.76 [1.04-7.33], ptrend=0.03). Higher IGF-I was associated with a non-significant decrease in risk for invasive (ORT3 vs. T1: 0.79 [0.62-1.02], ptrend=0.07) and endometrioid tumors (ORT3 vs. T1: 0.55 [0.28-1.07], ptrend=0.07). The inverse association between IGF-I levels and risk of invasive EOC was stronger in analyses limited to women aged &lt;55 years at diagnosis (ORT3 vs. T1: 0.74 [0.57-0.96], ptrend=0.03). No associations were observed between pre-diagnostic progesterone, SHBG, placental GH, and AMH with EOC risk overall, or by tumor invasiveness and histology. The longitudinal study showed that hormone concentrations were more strongly correlated between consecutive trimesters of a pregnancy than between the 1st and 3rd trimesters. Further, 3rd trimester hormone concentrations can be estimated from 1st or 2nd trimester measurements. Conclusion: Higher pre-diagnostic androgens, estradiol, and IGF-I are associated with EOC risk, and associations differ by tumor invasiveness and histology.