Low load resistance training (LLRT) with blood flow restriction (BFR) is a novel form of exercise that has been demonstrated to increase muscle mass and strength. Combined with the fact that as individuals age they lose both of these parameters, LLRT with BFR has been put forward as a method to help reverse/prevent the associated sarcopenia of ageing. This research investigated the effect the effect of LLRT with BFR on muscle strength firstly in younger people and then an older population group following 4 weeks of training. Muscle function measurements of young and old people included dynamic strength, identified as one repetition maximum (1 RM), isometric strength and isokinetic torque at a range of velocities (0.52 2.09 rad.s-1). Vascular adaptations were also measured using venous occlusion plethysmography to assess rest blood flow (Rbf) and post occlusive reactive hyperemia (PObf). The mechanisms behind any adaptations were measured following acute responses of plasma hormones and growth factors (cortisol, growth hormone (GH), insulin like growth factor 1 (IGF-1), interleukin 6 (IL-6) and vascular endothelial growth factor (VEGF)) as well as local skeletal muscle gene expression (IGF-1Ea and MGF mRNA) to LLRT with BFR. LLRT with BFR increased (P < 0.05) all measurements of muscle strength by 13 30% in both young and older people. PObf was also increased (P < 0.05) following 4 weeks of LLRT with BFR in both population groups. Acute responses to LLRT with BFR identified an increase (P < 0.05) in GH and VEGF in older people. These are similar response to those seen in the young. Finally local gene expression of MGF mRNA was elevated (P < 0.05) 24 hours post LLRT with BFR in both young and older people. Any changes in muscle and blood flow adaptations may be as a result of increased hormones and growth factors at a circulation and local level. Key words: Blood flow restriction, blood flow, muscle strength, growth hormone, IGF-1

Foram utilizados 60 animais da raça Holandesa, distribuídos em 2 grupos de acordo com número de lactação e em dois tempos de acordo com época de concepção. Para Alu-1, animais com genótipo Leu/Leu apresentaram maior produção de leite na lactação (p = 0,016) e tendência para maior produção ao pico (p = 0,054) que animais do genótipo Leu/Val. Não foi possível demonstrar efeito de genótipo para Dde-1 sobre nenhuma das variáveis estudadas. Houve correlação positiva entre níveis de IGF-1 e P4 (R2 = 0,28 e p = 0,011), correlação negativa entre IGF-1 DEAs, produção na lactação e produção ao pico de lactação (R2=- 0,267 e p = 0,016; R2=- 0,424 e p =0,0001; R2= -0,231 e p=0,0001, respectivamente). Houve tendência de correlação positiva entre escore corporal e níveis de P4 (R2 = 0,216 e p = 0,053). Concluí-se que o polimorfismo do fragmento de restrição identificado pela Alu-1 possa ser utilizado como marcador genético importante para seleção de gado leiteiro, e que o polimorfismo para Dde-1 não se apresentou como um bom marcador genético a ser utilizado para a raça Holandesa, o nível plasmático de IGF-1 parece ser um bom indicador do retorno a atividade reprodutiva. / Sexty female Holstein cows were used in this experiment, animals were arranged random in two groups in agreement lactation number and conception time.Animals with ALU-1 genotipe showed biggest milk production in lactation (p = 0,016) and tendence to biggest milk peak production than Leu/Val animals. Dde-1 didn?t showed genotipe effect for any variavels. There were positive correlactin among IGF-1 and progesterone (R2 =0,280 e p = 0,011). There were negative correlactin among IGF-1 and days open, lactation production, peak lactation (R2=- 0,267 p = 0,016; R2=- 0,424 p =0,0001; R2= -0,231 p=0,0001, respective). There were tendence of positive correlaction (R2 = 0,216 p=0,053) among body condition and progesterone levels. Basead on the results of the experiment, the Alu-1 polymorphism could be used as genetic markers to milk cow selection; DDE-1 polymorphism isnt indicate like genetic marker in Holstein cows; the IGF-1 levels seens a good indicator to return reproduction activite.

Absence of functional phenylalanine hydroxylase results in phenylketonuria (PKU). Viable treatments remain few, expensive and secondary conditions such as osteopenia occur in most PKU patients. Objective 1: Given the recently described roles of gut microbes to aid host digestion, an orally administered genetically engineered probiotic as the delivery vehicle for enzyme replacement therapy was created. The engineered probiotic, pHENOMMenal, produced phenylalanine ammonia lyase with significant production of trans-cinnamate (phenylalanine cleavage product) in vitro and resulted in a reduction of 515 μM in blood phenylalanine when fed to PKU animals for 14 days (from 2307µM ± 264µM to 1792µM ± 261µM, n = 6, P < 0.05). The control probiotic produced no change in blood phenylalanine. Thus, pHENOMMenal treatment in PKU mice demonstrated engineered microbes could compensate for a metabolic deficiency of the host. Objective 2: Evaluate the PAHenu2 mouse model of PKU for a genetic discrepancy causing ocular enlargement and delayed development observed only after the PAHenu2 mutation was crossed to the C57BL/6J mouse. When compared to healthy littermates, ELISA indicated a consistent but insignificant decrease in plasma IGF-1 and an increase in ocular IGF-1 in PKU animals. SNP screening demonstrated a differential inheritance of IGF-1 alleles in healthy and PKU animals based on PAH allele inheritance. Ocular and developmental phenotypes in the PAHenu2 colony match those described in previous IGF-1 studies. Understanding the IGF-1 inheritance discrepancy will enable better osteopenia research using PAHenu2 mice and allow breeding of a healthier mouse colony for continued research. Collectively the results from this work describe a new therapeutic approach for treatment of PKU as well as a better understanding of the PAHenu2 mouse model to study this disease.

O transtorno bipolar (TB) é uma doença crônica, altamente incapacitante e sua fisiopatologia não esta bem esclarecida. Apresenta altas taxas de comorbidades clínicas e risco de suicídio trazendo prejuízos e custos significativos para o indivíduo com a doença e para a sociedade. Existem evidências que relacionam o TB à alterações no fator de crescimento semelhante à insulina tipo 1 (IGF- 1) e nos sistemas endócrino e imune. O objetivo deste estudo foi avaliar os níveis séricos de IGF-1 em pacientes bipolares comparados com indivíduos controle e sua relação com a inflamação. Foram selecionados 31 pacientes com TB e 33 controles saudáveis. Foram avaliadas as concentrações séricas de IGF-1, hormônio do crescimento (GH), insulina e fator de necrose tumoral α (TNF-α). Como resultado deste estudo, observamos que os níveis séricos de IGF-1 estavam aumentados em pacientes com TB em relação aos controles (p = 0,001). Não foram observadas diferenças estatisticamente significativas entre os grupos nas dosagens de insulina, GH e TNF- α. Este estudo sugere uma associação entre IGF-1 na fisiopatologia do transtorno bipolar. É possível que este aumento periférico esteja relacionado com um aumento da resistência do IGF- 1 no SNC, reduzindo assim a sua ação neuroprotetora. / Bipolar disorder (BD) is a chronic, highly debilitating and its pathophysiology is not well understood. It offers high rates of clinical comorbidities and suicide risk causing losses and significant costs to the individual with the disease and society. There is evidence that relates to changes in TB-like growth factor type 1 insulin (IGF-1) and the endocrine and immune systems. The aim of this study was to evaluate serum levels of IGF-1 in bipolar patients compared with control subjects and their relationship to inflammation. We selected 31 patients with TB and 33 healthy controls. Serum concentrations of IGF-1, growth hormone, were evaluated (GH), insulin and tumor necrosis factor α (TNF-α). As a result of this study, we observed that serum levels of IGF-1 were increased in TB patients compared to controls (p = 0.001). No statistically significant differences were found between groups in insulin dosages, GH, and TNF-α. This study suggests an association between IGF-1 in the pathophysiology of bipolar disorder. It is possible that this increase is associated with peripheral increased IGF-1 resistance in the CNS, thus reducing its neuroprotective action.

O objetivo deste estudo foi analisar as concentrações do fator de crescimento semelhante à insulina tipo 1 (IGF1) e insulina (INS) no fluido folicular durante foliculogênese em éguas, e localizar o receptor tipo 1 de IGF (IGF1R) nas paredes foliculares. Durante a estação reprodutiva, 40 éguas mestiças enviadas para abate em matadouro, com idades variando entre 6 e 16 anos foram utilizadas. As éguas foram abatidas de forma humanitária. Os tratos reprodutivos internos foram recuperados dentro de 10 min após o abate, ovários de éguas cíclicas foram separados do resto do aparelho. As éguas foram classificadas em quatro grupos: G1 – Folículo ≤ 13,5 mm e CL identificável; G2 – Folículo de 13,6 - 22,5 mm e CL identificável; G3 – Folículo de 22,6 - 31,5 mm e CL identificável; G4 – Folículo ≥ 31,6 mm e CL difícil de identificar. O fluido folicular (FF) foi aspirado e armazenado a -80 ° C até a sua utilização. Após a punção do FF, um fragmento da porção ventral da parede folicular foi removido para realizar a imunohistoquímica para localização de receptores IGF1R. O IGF1 livre foi determinado por ensaio imunoradiométrico. A INS foi determinada por um radioimunoensaio de fase líquida. A concentração de IGF1 livre e INS no FF aumentou com o crescimento folicular (P < 0,05). O escore imunohistoquímico de IGF1R nas células da granulosa e da teca interna da parede folicular equina em diferentes grupos aumentou (P < 0,05) com aumento folicular. Concluímos que as concentrações de IGF1 livre e INS no FF e de IGF1R em paredes foliculares de éguas aumentam com o crescimento folicular durante a foliculogênese, dando evidência do papel importante do IGF1 e INS no desenvolvimento folicular, e uma interação entre ambos hormônios na fisiologia reprodutiva ovariana. Sendo a égua um modelo de pesquisa para estudos comparativos na dinâmica folicular para consideração em mulheres, a conclusão deste estudo pode ser aplicada, também, a humanos. / The aim of this study was to analyze free type 1 insulin-like growth factor (IGF1) and insulin (INS) concentrations in follicular fluid during folliculogenesis in mares, and to localize type 1 IGF receptor (IGF1R) on follicular walls. During the breeding season, 40 mixed-breed mares sent to slaughter at an abattoir, with ages ranging between 6 to 16 years were used. Mares were humanely slaughtered. Internal reproductive tracts were recovered within 10 min after slaughter; ovaries of cyclic mares were separated from the rest of the tract. Mares were categorized into four groups: G1 – Follicle ≤ 13.5 mm and CL identifiable; G2 – Follicle of 13.6 to 22.5 mm and CL identifiable; G3 – Follicle of 22.6 to 31.5 mm and CL identifiable; G4 – Follicle ≥ 31.6 mm and CL difficult to identify. The follicular fluid (FF) was aspirated and stored at -80°C until use. After puncture of the FF, a fragment of the ventral portion of the follicular wall was removed to perform immunohistochemistry to localize IGF1R receptors. Free IGF1 was determined by immunoradiometric assay. INS was determined by a liquid phase radioimmunoassay. The concentration of free IGF1 and INS in FF increased with follicle growth (P < 0.05). Immunohistochemical score of IGF1R on granulosa and internal theca cells from equine follicular wall on different groups increase (P < 0.05) with follicular raise. We conclude that free IGF1 and INS concentrations in FF and IGF1R in follicular walls of mares increase with follicular growth during folliculogenesis, giving evidence of important role of IGF1 and INS in follicular development, and an interaction between both hormones in ovarian reproductive physiology. As the mare is a model research for comparative studies in follicular dynamics for consideration in women, the conclusion of this study can be applied to humans also.

Foram utilizados 60 animais da raça Holandesa, distribuídos em 2 grupos de acordo com número de lactação e em dois tempos de acordo com época de concepção. Para Alu-1, animais com genótipo Leu/Leu apresentaram maior produção de leite na lactação (p = 0,016) e tendência para maior produção ao pico (p = 0,054) que animais do genótipo Leu/Val. Não foi possível demonstrar efeito de genótipo para Dde-1 sobre nenhuma das variáveis estudadas. Houve correlação positiva entre níveis de IGF-1 e P4 (R2 = 0,28 e p = 0,011), correlação negativa entre IGF-1 DEAs, produção na lactação e produção ao pico de lactação (R2=- 0,267 e p = 0,016; R2=- 0,424 e p =0,0001; R2= -0,231 e p=0,0001, respectivamente). Houve tendência de correlação positiva entre escore corporal e níveis de P4 (R2 = 0,216 e p = 0,053). Concluí-se que o polimorfismo do fragmento de restrição identificado pela Alu-1 possa ser utilizado como marcador genético importante para seleção de gado leiteiro, e que o polimorfismo para Dde-1 não se apresentou como um bom marcador genético a ser utilizado para a raça Holandesa, o nível plasmático de IGF-1 parece ser um bom indicador do retorno a atividade reprodutiva. / Sexty female Holstein cows were used in this experiment, animals were arranged random in two groups in agreement lactation number and conception time.Animals with ALU-1 genotipe showed biggest milk production in lactation (p = 0,016) and tendence to biggest milk peak production than Leu/Val animals. Dde-1 didn?t showed genotipe effect for any variavels. There were positive correlactin among IGF-1 and progesterone (R2 =0,280 e p = 0,011). There were negative correlactin among IGF-1 and days open, lactation production, peak lactation (R2=- 0,267 p = 0,016; R2=- 0,424 p =0,0001; R2= -0,231 p=0,0001, respective). There were tendence of positive correlaction (R2 = 0,216 p=0,053) among body condition and progesterone levels. Basead on the results of the experiment, the Alu-1 polymorphism could be used as genetic markers to milk cow selection; DDE-1 polymorphism isnt indicate like genetic marker in Holstein cows; the IGF-1 levels seens a good indicator to return reproduction activite.

<p>Multiple myeloma (MM) is a hematological tumor of plasma blast/plasma cell origin heterogeneous with respect to the morphological differentiation stage of the tumor cells, genetic alterations and course of disease. A challenge in MM research is to overcome resistance to therapy, which inevitably arises. In this thesis, we have used different strategies to sensitize MM cells to apoptosis and explored possible mechanisms of apoptotic control by the insulin-like growth factor-1 receptor (IGF-1R) survival pathway.</p><p>mTOR is a key molecule in the regulation of translation activated by survival signaling pathways in MM. We demonstrate that the mTOR-inhibitor rapamycin alone induced apoptosis in primary MM cells. In addition, rapamycin sensitized MM cells to apoptosis induced by dexamethasone, a glucocorticoid frequently used in MM therapy. MM survival factors IGF-1 and IL-6 could neither restore phosphorylation of the mTOR target p70S6K, nor cell growth inhibited by rapamycin and dexamethasone.</p><p>To study the regulation of inhibitors of apoptosis (IAP), we induced apoptosis and cell cycle arrest with dexamethasone and simultaneously abrogated IGF-1R signaling using the antagonistic antibody αIR3 or the selective IGF-1R inhibitor picropodophyllin (PPP). Dexamethasone transiently up-regulated c-IAP2. The subsequent down-regulation of c-IAP2 and XIAP was associated with the onset of apoptosis. c-IAP2 and XIAP levels further decreased when enhancing dexamethasone-induced apoptosis using αIR3 or PPP indicating a role for IAPs in regulating resistance to apoptosis in MM.</p><p>Finally, we explored glycogen synthase kinase (GSK)3 as a possible pro-apoptotic molecule and its role in regulating sensitization to apoptosis. We show that inhibition of GSK3 counteracts growth inhibition induced by dexamethasone alone and in combinatorial treatments with inhibitors against PI 3-kinase, mitogen-activated protein kinase (MEK), mTOR and IGF-1R. CT99021 also reversed cell cycle arrest induced by LY294002 or rapamycin. Importantly, the GSK3 inhibitor CT99021 sustained viability in untreated and dexamethasone-treated primary MM cells.</p>

Multiple myeloma (MM) is a hematological tumor of plasma blast/plasma cell origin heterogeneous with respect to the morphological differentiation stage of the tumor cells, genetic alterations and course of disease. A challenge in MM research is to overcome resistance to therapy, which inevitably arises. In this thesis, we have used different strategies to sensitize MM cells to apoptosis and explored possible mechanisms of apoptotic control by the insulin-like growth factor-1 receptor (IGF-1R) survival pathway. mTOR is a key molecule in the regulation of translation activated by survival signaling pathways in MM. We demonstrate that the mTOR-inhibitor rapamycin alone induced apoptosis in primary MM cells. In addition, rapamycin sensitized MM cells to apoptosis induced by dexamethasone, a glucocorticoid frequently used in MM therapy. MM survival factors IGF-1 and IL-6 could neither restore phosphorylation of the mTOR target p70S6K, nor cell growth inhibited by rapamycin and dexamethasone. To study the regulation of inhibitors of apoptosis (IAP), we induced apoptosis and cell cycle arrest with dexamethasone and simultaneously abrogated IGF-1R signaling using the antagonistic antibody αIR3 or the selective IGF-1R inhibitor picropodophyllin (PPP). Dexamethasone transiently up-regulated c-IAP2. The subsequent down-regulation of c-IAP2 and XIAP was associated with the onset of apoptosis. c-IAP2 and XIAP levels further decreased when enhancing dexamethasone-induced apoptosis using αIR3 or PPP indicating a role for IAPs in regulating resistance to apoptosis in MM. Finally, we explored glycogen synthase kinase (GSK)3 as a possible pro-apoptotic molecule and its role in regulating sensitization to apoptosis. We show that inhibition of GSK3 counteracts growth inhibition induced by dexamethasone alone and in combinatorial treatments with inhibitors against PI 3-kinase, mitogen-activated protein kinase (MEK), mTOR and IGF-1R. CT99021 also reversed cell cycle arrest induced by LY294002 or rapamycin. Importantly, the GSK3 inhibitor CT99021 sustained viability in untreated and dexamethasone-treated primary MM cells.

Wir untersuchten Liquor- und Serumproben von 40 an einem shuntversorgten Hydrocephalus erkrankten Kindern auf die Konzentration der Zytokine bFGF, TGF-β1, VEGF, IL-6, IGF-1 und Leptin sowie deren Korrelation mit dem Risiko von Shuntinsuffizienzen. Dabei konnten wir die Hypothese bestätigen, dass erhöhte Konzentration der fibrogenen Zytokine bFGF und TGF-β1 im Serum bzw. Liquor mit einem erhöhten Risiko für operationspflichtige Shuntinsuffizienzen durch Obstruktion des Schlauchsystems einhergehen, und dass diese Komplikationen mit steigenden Zytokinkonzentrationen umso eher eintreten. Außerdem war bFGF im Liquor von Kindern, die zum Abnahmezeitpunkt an einer Shuntdysfunktion durch Obstruktion oder Einwachsen des Shunts litten, signifikant höher als bei Kindern, die zum Zeitpunkt der Abnahme keine Shuntdysfunktion aus eben genannten Gründen hatten. Des Weiteren fanden wir Konzentrationsunterschiede für IL-6 im Liquor zwischen den einzelnen Ursachen der Erkrankung, wobei das Zytokin am höchsten bei Tumorpatienten war, gefolgt von posthämorrhagischem und postmeningitischem Hydrocephalus, und am niedrigsten bei Kindern mit kongenitaler ZNS-Fehlbildung.

The purpose of this study was to look at the genotoxic and cytotoxic effects of benzo-á-pyrene (BáP), a chemical mutagen that is present in cigarette smoke, on the insulin-like growth factor-I (IGF-I) gene. Women who smoke during pregnancy are more likely to have a growth-restricted baby. We hypothesized that BáP exerts its effects through genotoxic and cytotoxic avenues. The cytotoxicity is manifested by chromosomal abnormalities and a decrease in the rate of cell division. The genotoxicity is manifested by changes in certain genes known to be important in mammalian fetal development such as IGF-I. IGF-I is implicated in intrauterine growth restriction (IUGR), a problem that greatly increases the risk of perinatal morbidity and mortality. To futher understand the mechanism by which BáP influences the normal growth and development of human placental cells, human placental trophoblast cells from an established immortalized cell line were utilized. Cells were cultured in appropriate media, starved (using starvation "Serum Free Medium"), and treated with two doses of BáP, 1µM (dose 1) and 5µM (dose 2). Chromosomes were prepared for cytogenetic analysis and visualized using light microscopy after Giemsa staining. Chromosomal aberrations were identified and the rate of cell division was determined through the analysis of the mitotic index for treated cells compared to a control group. To further understand the influence of BáP on the IGF-I gene expression level, RNA was extracted from control and treated cells, from which cDNA was synthesized and used for further analysis using polymerized chain reaction (PCR). The PCR results were used to better understand the genotoxicity of BáP, while chromosomal aberration analysis was used to determine the cytotoxic effects of BáP on human placental cells. Our results indicate that many chromosomal abnormalities were present in the treated groups compared to the control group. In addition, there was a significant decrease in the mitotic index of the BáP-treated cells (MI=0.3%) verses the control group (MI=0.93%), p value 0.0447. Through the PCR assay, we speculate that there is a dose-related response to BáP of the IGF-I RNA expression level, with low levels in the treated groups compared to the control group. We conclude from these results that BáP influences placental cells at both the gene and chromosome level. It also affects the cell cycle of human placental cells. It is known that smoking is deleterious for fetal development. We believe that the current study brings us closer to understanding the mechanism by which smoking can lead to fetal growth restriction.