Introdução: Não foram encontrados na literatura dados sobre a prevalência de diabetes mellitus em mães de crianças com fissura labiopalatina. Dada a relevância do tema esse foi o principal objetivo da presente investigação. Método: Após aprovação do comitê de ética e pesquisa e obtenção do consentimento livre e esclarecido, foram entrevistadas 325 mulheres, mães biológicas de crianças com fissuras labiopalatinas com idades de 0 a 3 anos matriculadas no Hospital de Reabilitação de Anomalias Craniofaciais da Universidade de São Paulo, HRAC-USP. Foi aplicado questionário com 24 questões onde obteve-se a identificação das mães, o tipo de diabetes, os principais sinais clínicos, as comorbidades associadas, hipertensão, obesidade, medicamentos e drogas lícitas e ilícitas usadas durante a gravidez. Também foram aferidos a pressão arterial e o perímetro abdominal das mães e anotados os valores da glicemia de jejum na primeira consulta pré-natal. Resultados: Os resultados obtidos mostraram que 88 mulheres apresentavam diabetes mellitus, sendo 78 com diabetes mellitus gestacional, 05 com diabetes mellitus tipo 1 e 05 com diabetes mellitus tipo 2. A prevalência de diabetes mellitus em mães de crianças com fissuras labiopalatinas foi de 27,08%, comparativamente maior (p<0,01) aos valores encontrados na população adulta brasileira (7,6%). Dessas 88 mulheres, foram excluídos os fatores que de alguma forma pudessem influenciar o aparecimento de fissuras labiopalatinas e obteve-se o percentual de 16%, onde a hiperglicemia materna nessas mães foi o único fator provável no aparecimento dessas fissuras. Conclusão: A prevalência de diabetes mellitus em mães de crianças com fissuras labiopalatinas foi de 27,08%. (p<0,01). Extraídos os fatores tidos como confundidores, tais como medicamentos usados na gestação, álcool, tabaco, drogas ilícitas, obesidade e hipertensão arterial, restaram 52 mães que tinham somente a hiperglicemia materna como fator isolado, num total de 16%. / Introduction: there was no data found on the literature regarding the prevalence of diabetes mellitus in mothers of children born with cleft lip and palate. Given the relevance of the subject, that was the main objective of this investigation. Method: after approval by the research and ethics committee, and the free, informed consent firmed by the mothers, interviews were conducted with 325 women biological mothers of children aged between 0 to 3 years, born with cleft lip and palate, registered at Hospital for Rehabilitation of Craniofacial Anomalies, University of São Paulo, HRAC-USP. A 24-question survey was applied to collect data regarding the mothers identification, type of diabetes, clinical data, associated comorbidities, hypertension, obesity, medication and legal or illegal drugs used during pregnancy. Blood pressure and abdominal circumference were measured, and fasting blood glucose levels measured at the first prenatal visit. Results: It was found that 88 women had diabetes mellitus, 78 of which had gestational diabetes, 05 had type 1 diabetes and 05 had type 2 diabetes the prevalence of diabetes in mothers of children with cleft lip and palate was 27.08%, a number significantly higher (p<0.01) than the rates found among the Brazilian adult population (7.6%). From this group of 88 women we have suppressed the factors that could contribute to the occurrence of cleft lip and palate in some way, obtaining a prevalence of 16% in which maternal hyperglycemia was the only likely cause of cleft lip and palate. Conclusion: The prevalence of diabetes mellitus in mothers of children born with cleft lip and palate was 27.08% (p<0.01). Suppressing other potentially confusing factors like medication used during pregnancy, alcohol, tobacco, illegal drugs, obesity and hypertension, there were 52 mothers who had only maternal hyperglycemia as an isolated factor, representing 16% of the total studied population.

Diabetes Mellitus (DM) é uma síndrome metabólica complexa, causada pela secreção diminuída ou ausente de insulina pelas células beta pancreáticas, levando a hiperglicemia. A hiperglicemia promove a glicação de proteínas e, conseqüentemente, o aparecimento de produtos finais da glicação avançada (AGEs). Atualmente, os pacientes diabéticos são monitorados pela determinação dos níveis de glicemia e hemoglobina glicada (HbA1c). As complicações geradas pela hiperglicemia podem ser divididas em micro e macrovasculares, representadas por retinopatias, nefropatias, neuropatias e doenças cardiovasculares. A albumina (HSA) é a proteína sérica mais abundante no organismo humano e está sujeita à glicação. A protoporfirina XI (PpIX) é a molécula precursora da síntese do heme, componente estrutural da hemoglobina. Ensaios in vitro e em animais indicaram que a hiperglicemia promove uma diminuição de sua concentração em eritrócitos. A espectroscopia de fluorescência é uma técnica bastante utilizada na área biomédica. A autofluorescência corresponde à fluorescência intrínseca presente em algumas moléculas, estando esta associada à estrutura das mesmas. O objetivo deste trabalho foi utilizar a técnica de espectroscopia de fluorescência para mensurar os níveis de autofluorescência da PpIX eritrocitária e AGE-HSA em pacientes diabéticos e indivíduos saudáveis e compará-los com os níveis de glicemia e HbA1c. Este estudo foi realizado com 151 indivíduos (58 controles e 93 diabéticos). Os dados epidemiológicos de pacientes e controles foram obtidos nos prontuários médicos. Para os indivíduos controle, os valores de glicemia foram adquiridos dos prontuários médicos e os níveis de Hb1Ac obtidos pela utilização de kits comerciais. A determinação da autofluorescência da PpIX foi realizada com excitação de 405 nm e emissão de 632 nm. Para a determinação do AGE-HSA foi realizada excitação de 370 nm e emissão de 455 nm. Aproximadamente 50% dos diabéticos apresentaram lesões micro ou macrovasculares decorrentes da hiperglicemia. Não foram observadas diferenças significativas nos valores de intensidade de emissão de PpIX entre os grupos estudados (P=0,89). Na análise do AGE-HSA observou-se diferenças significativas dos valores de intensidade de emissão entre os dois grupos, sendo este valor 1,45 vezes maior para o grupo de indivíduos diabéticos (P<0,0001). Os pacientes com complicações diabéticas apresentavam intensidade de emissão de fluorescência 1,19 vezes maior que os indivíduos sem complicações decorrentes da doença (P= 0,01), mesmo não havendo diferenças significativas nos valores de HbA1c entre os dois grupos. Concluímos que a espectroscopia de fluorescência foi uma técnica eficaz na identificação da autofluorescência da PpIX e do AGE-HSA. A PpIX não foi um biomarcador eficiente para o acompanhamento do DM. A determinação dos níveis de autofluorescência do AGE-HSA foi eficiente para a discriminação entre os grupos e para o monitoramento da progressão da doença, podendo ser mais eficiente que a dosagem de HbA1c. A espectroscopia de fluorescência é uma técnica simples, rápida e de baixo custo para o acompanhamento de indivíduos diabéticos. / Diabetes Mellitus (DM) comprises a complex metabolic syndrome, caused by reduced or absent secretion of insulin by pancreatic beta cells, leading to hyperglycemia. Hyperglycemia promotes glycation of proteins and, consequently, the appearance of advanced glycation end products (AGEs). Currently, diabetic patients are monitored by determining levels of glucose and glycated hemoglobin (HbA1c). The complications caused by hyperglycemia may be divided into micro and macrovascular complications, represented by retinopathy, nephropathy, neuropathy and cardiovascular disease. Albumin (HSA) is the most abundant serum protein in the human body and is subject to glycation. The Protoporphyrin IX (PpIX) is the precursor molecule of heme synthesis, structural component of hemoglobin. The in vitro and animals studies have indicated that hyperglycemia promotes a decrease in its concentration in erythrocytes. The fluorescence spectroscopy is a technique widely used in biomedical field. The autofluorescence corresponds to the intrinsic fluorescence present in some molecules, this being associated with the same structure. The aim of this study was to use fluorescence spectroscopy to measure levels of erythrocyte PpIX autofluorescence and AGE-HSA in diabetic and healthy subjects and compare them with levels of blood glucose and HbA1c. This study was conducted with 151 subjects (58 controls and 93 diabetics). Epidemiological data of patients and controls were obtained from medical records. For control subjects, blood glucose levels were obtained from medical records and levels of Hb1Ac obtained by using commercial kits. The determination of the PpIX autofluorescence was performed with excitation at 405 nm and emission at 632 nm. Determination of AGE-HSA was performed with excitation at 370 nm and emission at 455 nm. Approximately 50% of diabetic had micro and macrovascular lesions resulting from hyperglycemia. There were no significant differences in the PpIX emission intensity values between groups (P = 0.89). In the analysis of AGE-HSA was observed significant differences in the values of emission intensity between the two groups, and this value was 1.45-fold greater for the group of diabetic (P <0.0001). Patients with diabetic complications had fluorescence emission intensity of 1.19-fold higher than individuals without disease complications (P = 0.01), even with no significant differences in HbA1c values between the two groups. We conclude that fluorescence spectroscopy was an effective technique in the identification of the PpIX autofluorescence and AGE-HSA. The PpIX was not an effective biomarker for the monitoring of diabetes. The determination of AGE-HSA autofluorecência was efficient for the discrimination between groups and monitoring disease progression, may be more effective than HbA1c dosage. The fluorescence spectroscopy is a simple, fast and low cost for the monitoring of diabetic patients.

研究介紹：炎症與纖維化是糖尿病腎病（DN）的主要特徵。研究發現生長轉化因子TGF-β/Smad信號在糖尿病所致炎症與纖維化中均起重要作用。我們認為TGF-β/Smad信號通路失調是導致糖尿病腎損傷的主要機制，恢復信號通路或有治療價值。為此我們通過以下研究證實：（1）研究Smad7基因在DN中的作用，及評估Smad7基因治療效果；（2）研究miR-29在DN中的作用，及評估miR-29基因治療效果；（3）研究C反應蛋白（CRP）在DN中的作用及機制。 / 研究方法：（1）利用Smad7基因敲除（KO）小鼠建立糖尿病小鼠，並研究Smad7基因在DN的作用，並在链脲佐菌素（STZ）誘導的糖尿病大鼠上利用微泡導入Smad7基因治療觀察其療效；（2）在10週齡db/db小鼠上利用微泡導入可誘導的miR-29b基因，觀察miR-29b在糖尿病腎病中的作用，並用miR-29敲除或高表達細胞株研究其機制；（3）利用CRP轉基因小鼠誘導糖尿病，觀察CRP在DN中的作用，及以高糖和/或CRP刺激腎小管細胞研究CRP的致病機制。 / 研究結果：我們發現（1）糖尿病Smad7 KO小鼠出現更嚴重的腎損傷，包括蛋白尿增加，腎臟炎症及纖維化加重。進一步研究發現Smad7下調所致TGF-β/Smad和NF-kB信號過度活化是導致腎臟炎症及纖維化加重的重要原因。運用基因治療恢復糖尿病大鼠的Smad7水平，發現能夠減輕蛋白尿增加，及抑制TGF-β/Smad引起的纖維化和NF-kB所致炎症反應；（2）我們發現miR-29b在20週齡db/db小鼠比10週齡的顯著降低，並伴隨有蛋白尿加重，腎臟纖維化和炎症反應增加，及TGF-β/Smad，NF-kB，T-bet信號上調，而miR-29b基因治療能減輕蛋白尿，及減輕腎臟纖維化和炎症反應增加，及TGF-β/Smad，NF-kB，T-bet信號上調。體外實驗證實AGEs刺激miR-29敲除細胞株增加纖維化，伴隨有TGF-β/Smad3及炎症因子上調，而刺激高表達細胞株能抑制纖維化，及TGF-β/Smad和炎症因子下調；（3）糖尿病CRP轉基因小鼠出現更嚴重的腎損傷，出現蛋白尿和腎損傷分子-1上升、巨噬細胞和T細胞侵潤、炎症和纖維化增加，並伴有CRP受體（CD32a）上調、TGF-β/Smads及NFκB/p65信號過度活化。體外實驗進一步證實CRP通過其受體CD32a/CD64增加炎症和纖維化。另外證實CRP與高糖有協同作用。 / 結論：TGF-β/Smad信號通路是糖尿病腎病的重要致病機制。糖尿病腎病導致Smad7、miR-29b下調，運用基因治療恢復其表達能減輕糖尿病腎損傷。 / Diabetic nephropathy (DN) is characterized by renal fibrosis and inflammation. Increasing evidence shows that TGF-β/Smad signaling plays a critical role in DN. This thesis tested a hypothesis that TGF-β/Smad signaling may play a central role in diabetic kidney injury and targeting this pathway may represent a novel therapy for DN. The hypothesis was tested in a type-1 model of diabetes induced in Smad7 knockout (KO) or CRP transgene, and the therapeutic potential for DN was examined by overexpressing renal Smad7 or miR-29b in both type-1 or type-2 models of diabetes. / As described in Chapter Three, the protective role and therapeutic potential of Smad7 in diabetic kidney disease was investigated in streptozotocin-induced diabetic model in Smad7 KO mice and in diabetic rats given Smad7 gene transfer using an ultrasound-microbubble-mediated technique. Results showed that Smad7 KO mice developed more severe diabetic kidney injury than wild type (WT) mice as evidenced by a signicant increase in microalbuminuria, renal brosis, and renal inammation, which was mediated by enhanced activation of both TGF-β/Smads and NF-κB signaling pathways. To develop a therapeutic potential for diabetic kidney disease, Smad7 gene was transferred into the kidney, which results in high levels renal Smad7, thereby blocking microalbuminuria, TGF-β/Smad3-mediated renal brosis and NF-κB/p65-driven renal inammation in diabetic rats. / To test a novel hypothesis that TGF-β/Smad3-mediated DN via the Smad3-dependent miR-29, in Chapter Four, the role and mechanisms of miR-29b in DN were examined in vitro in a stable mesangial cell line with overexpression or knockdown of miR-29b and the therapeutic effect of miR-29b on DN was developed by delivering a Dox-inducible miR-29b into 10-week db/db mice. Results showed that addition of AGEs induced a loss of miR-29b with increased fibrosis and inflammation in mesangial cells, which was further enhanced with miR-29b knockdown, but inhibited by overexpressing miR-29b. In db/db mice, reduction of renal miR-29b over the 20 week time was associated with a marked increase in microabluminuria, renal fibrosis and inflammation. Restoring miR-29b resulted in inhibition of kidney injuries by blocking TGF-β/Smad3-mediated renal fibrosis, NF-kB/p65-driven renal inflammation, and importantly, the Th1-dependent immune response, revealing a critical role and therapeutic potential for miR-29b in the pathogenesis of DN. / Finally, diabetic kidney injury was also assessed in under high inflammation conditions in CRP transgenic (Tg) mice. As shown in Chapter Five, CRP Tg mice developed more severe diabetic kidney injury than WT mice, as evidenced by a significant increase in microalbuminuria and kidney injury molecule-1, macrophages and T cells, and upregulation of pro-inflammatory cytokines and extracellular matrix. CRP-mediated DN was associated with upregulation of CRP receptor, CD32a, and over-activation of the TGF-β/Smads and NFκB/p65 signaling pathways. These findings were further confirmed in vitro under high levels of CRP. In addition, CRP was induced by high glucose, which synergistically promoted high glucose-mediated renal inflammation and fibrosis, suggesting a positive feedback-loop between CRP and high glucose under diabetic conditions. / In conclusion, TGF-β/Smads play critical roles in the pathogenesis of DN. Loss of renal Smad7 and miR-29b may be a key mechanism of DN. Thus, over-expression of Smad7 or miR-29b may represent novel therapeutic strategies for diabetic kidney complications. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Chen, Haiyong. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 202-236). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / ABSTRACT --- p.ii / DECLARATION --- p.vi / ACKNOWLEDGEMENT --- p.vii / PUBLICATIONS --- p.ix / PRESENTATIONS/AWARDS --- p.xi / TABLE OF CONTENTS --- p.xii / LIST OF ABBREVIATIONS --- p.xxii / LIST OF FIGURES/TABLES --- p.xxiv / Chapter CHAPTER ONE --- INTRODUCTION --- p.1 / Chapter 1.1 --- TGF-β superfamily --- p.2 / Chapter 1.2 --- TGF-β/Smad signaling pathway --- p.3 / Chapter 1.2.1 --- TGF-β --- p.3 / Chapter 1.2.1.1 --- TGF-β structure --- p.3 / Chapter 1.2.1.2 --- Activation of latent TGF-β --- p.4 / Chapter 1.2.1.3 --- Latent TGF-β receptors --- p.6 / Chapter 1.2.2 --- TGF-β signaling pathway --- p.7 / Chapter 1.2.2.1 --- Receptors --- p.7 / Chapter 1.2.2.2 --- Smads --- p.10 / Chapter 1.2.2.3 --- Smad-dependent TGF-β signaling pathways --- p.13 / Chapter 1.2.2.4 --- Smad-independent TGF-β signaling pathways --- p.14 / Chapter 1.3 --- Diabetes nephropathy --- p.15 / Chapter 1.3.1 --- Diabetes Mellitus --- p.15 / Chapter 1.3.2 --- Type 1 and type 2 diabetes --- p.16 / Chapter 1.3.3 --- Diabetic complications --- p.16 / Chapter 1.3.4 --- Cellular and molecular mechanisms in diabetic complications --- p.17 / Chapter 1.3.4.1 --- Increased polyol pathway flux --- p.17 / Chapter 1.3.4.2 --- Increased advanced glycation end-products (AGEs) formation --- p.18 / Chapter 1.3.4.3 --- Activation of protein kinase C (PKC) isoforms --- p.20 / Chapter 1.3.4.4 --- Increased hexosamine pathway flux --- p.22 / Chapter 1.3.4.5 --- Increased Reactive Oxygen Species --- p.23 / Chapter 1.3.5 --- Diabetic kidney injuries --- p.24 / Chapter 1.3.5.1 --- Exacerbation of renal structure and function --- p.24 / Chapter 1.3.5.2 --- Fibrosis in diabetic nephropathy --- p.25 / Chapter 1.3.5.3 --- Inflammation in diabetic nephropathy --- p.26 / Chapter 1.4 --- Role of TGF-β/Smad signaling pathway in diabetic nephropathy --- p.28 / Chapter 1.4.1 --- Smad-depedent signaling in diabetic nephropathy --- p.28 / Chapter 1.4.2 --- Cross talk between Smads and other signaling pathways in diabetic nephropathy --- p.30 / Chapter 1.4.3 --- TGF-β/Smads and MicroRNA regulation in diabetic nephropathy --- p.32 / Chapter CHAPTER TWO --- MATERIALS AND METHODS --- p.35 / Chapter 2.1 --- Materials --- p.36 / Chapter 2.1.1 --- Regents and equipment --- p.36 / Chapter 2.1.1.1 --- Reagents and equipment for cell culture --- p.36 / Chapter 2.1.1.2 --- Reagents and equipment for real-time RT-PCR --- p.37 / Chapter 2.1.1.3 --- Reagents and equipment for western blotting --- p.38 / Chapter 2.1.1.4 --- Reagents and equipment for immunohistochemistry --- p.39 / Chapter 2.1.1.5 --- Reagents and equipment for in situ hybridization --- p.40 / Chapter 2.1.1.6 --- Reagents and equipment for plasmid purification --- p.40 / Chapter 2.1.1.7 --- Reagents and equipment for genotyping --- p.41 / Chapter 2.1.1.8 --- Other reagents --- p.41 / Chapter 2.1.2 --- Buffers --- p.42 / Chapter 2.1.2.1 --- Western blotting buffer --- p.42 / Chapter 2.1.2.2 --- Immunohistochemistry buffer --- p.45 / Chapter 2.1.2.3 --- ELISA buffers --- p.47 / Chapter 2.1.2.4 --- In Situ hybridization buffer --- p.48 / Chapter 2.2.2 --- Antibodies --- p.49 / Chapter 2.2.3 --- Primer sequences --- p.49 / Chapter 2.2 --- Methods --- p.56 / Chapter 2.2.1 --- Animal model --- p.56 / Chapter 2.2.1.1 --- Animals --- p.56 / Chapter 2.2.1.2 --- Diabetic animal models --- p.57 / Chapter 2.2.2 --- Sample Collection --- p.59 / Chapter 2.2.2.1 --- Urine collection --- p.59 / Chapter 2.2.2.2 --- Plasma collection --- p.59 / Chapter 2.2.2.3 --- Tissue collection --- p.60 / Chapter 2.2.2.4 --- Paraffin embedding --- p.60 / Chapter 2.2.3 --- Ultrasound-microbubble-mediated gene transfer system --- p.61 / Chapter 2.2.3.1 --- Smad7 gene therapy --- p.61 / Chapter 2.2.3.2 --- miR-29 gene therapy --- p.62 / Chapter 2.2.4 --- Microalbumin and renal function --- p.63 / Chapter 2.2.4.1 --- Microalbuminuria --- p.63 / Chapter 2.2.4.2 --- Creatinine measurement --- p.63 / Chapter 2.2.5 --- Enzyme-Linked Immunosorbent Assay (ELISA) --- p.64 / Chapter 2.2.6 --- Histology and immunohistochemistry --- p.64 / Chapter 2.2.6.1 --- Tissue process --- p.64 / Chapter 2.2.6.2 --- Periodic Acid-Schiff Staining (PAS) --- p.64 / Chapter 2.2.6.3. --- Immunohistochemistry (IHC) --- p.65 / Chapter 2.2.6.4 --- In Situ hybridization --- p.66 / Chapter 2.2.6.5 --- Quantitation of histology and IHC --- p.67 / Chapter 2.2.7 --- Cell culture --- p.67 / Chapter 2.2.8 --- Real-time polymerase chain reaction (PCR) --- p.69 / Chapter 2.2.9 --- Western Blotting --- p.70 / Chapter 2.3 --- Statistical analysis --- p.71 / Chapter CHAPTER THREE --- THE PROTECTIVE ROLE OF SMAD7 IN DIABETIC NEPHROPATHY --- p.72 / Chapter 3.1 --- Introduction --- p.73 / Chapter 3.2 --- Materials and methods --- p.74 / Chapter 3.2.1 --- Animal models --- p.74 / Chapter 3.2.2 --- Ultrasound-mediated gene transfer of inducible Smad7 gene-bearing microbubbles into the kidney --- p.74 / Chapter 3.2.3 --- Real-time PCR --- p.75 / Chapter 3.2.4 --- Western blotting --- p.75 / Chapter 3.2.5 --- Microalbuminuria and urinary creatinine analysis --- p.76 / Chapter 3.2.6 --- Histology and immunohistochemistry --- p.76 / Chapter 3.2.7 --- Statistical analysis --- p.77 / Chapter 3.3 --- Results --- p.77 / Chapter 3.3.1 --- Genotyping for Smad7 KO and WT mice --- p.77 / Chapter 3.3.2 --- Disruption of Smad7 enhances diabetic kidney injury --- p.78 / Chapter 3.3.3 --- Disruption of Smad7 enhanced fibrosis in diabetic kidney --- p.80 / Chapter 3.3.3.1 --- Collagen I is enhanced in diabetic Smad7 KO mice --- p.81 / Chapter 3.3.3.2 --- Collagen IV is enhanced in diabetic Smad7 KO mice --- p.82 / Chapter 3.3.3.3 --- Fibronectin is enhanced in diabetic Smad7 KO mice --- p.84 / Chapter 3.3.4 --- Disruption of Smad7 exacerbates inflammation in diabetic kidney --- p.85 / Chapter 3.3.4.1 --- Disruption of Smad7 increases IL-1β in diabetic kidney --- p.85 / Chapter 3.3.4.2 --- Disruption of Smad7 increases TNF-α in diabetic kidney --- p.86 / Chapter 3.3.4.3 --- Disruption of Smad7 Increases MCP-1 in diabetic kidney --- p.87 / Chapter 3.3.4.4 --- Disruption of Smad7 increases ICAM-1 in diabetic kidney --- p.88 / Chapter 3.3.4.5 --- Disruption of Smad7 increases macrophage infiltration in diabetic kidney --- p.90 / Chapter 3.3.5 --- Enhanced activation of TGF-β/Smad3 and NF-κB Signaling is a central mechanism by which disruption of Smad7 promotes diabetic renal fibrosis and inflammation --- p.91 / Chapter 3.3.5.1 --- Smad7 decreases in diabetic kidney --- p.91 / Chapter 3.3.5.2 --- Enhanced activation of TGF-β/Smad3 signaling pathway contributes to fibrosis in diabetic kidney --- p.92 / Chapter 3.3.5.3 --- Enhanced activation of NF-κB/p65 signaling pathway contributes to inflammation in diabetic kidney --- p.93 / Chapter 3.3.6 --- Smad7 transfection rate by gene therapy in diabetic rats --- p.94 / Chapter 3.3.7 --- Restoring Smad7 attenuates kidney injury in diabetic rats --- p.96 / Chapter 3.3.8 --- Restoring Smad7 attenuates renal fibrosis in diabetic rats --- p.98 / Chapter 3.3.8.1 --- Restoring Smad7 attenuates collagen I in diabetic kidney --- p.98 / Chapter 3.3.8.2 --- Restoring Smad7 attenuates collagen IV in diabetic kidney --- p.100 / Chapter 3.3.8.3 --- Restoring Smad7 attenuates collagen III in diabetic kidney --- p.101 / Chapter 3.3.9 --- Restoring Smad7 attenuates renal inflammation in diabetic rats --- p.104 / Chapter 3.3.9.1 --- Restoring Smad7 attenuates IL-1b in diabetic kidney --- p.104 / Chapter 3.3.9.2 --- Restoring Smad7 attenuates TNF-α in diabetic kidney --- p.106 / Chapter 3.3.9.3 --- Restoring Smad7 Attenuates MCP-1 in diabetic kidney --- p.107 / Chapter 3.3.9.4 --- Restoring Smad7 attenuates ICAM-1 in diabetic kidney --- p.109 / Chapter 3.3.9.5 --- Restoring Smad7 attenuates macrophage infiltration in diabetic kidney --- p.111 / Chapter 3.3.10 --- Blockade of activation of TGF-β/Smad3 and NF-κB signaling is a key mechanism by which overexpression of smad7 inhibits diabetic renal injury --- p.113 / Chapter 3.3.10.1 --- Restoring Smad7 inhibits activation of TGF-β/Smad3 signaling --- p.113 / Chapter 3.3.10.2 --- Restoring Smad7 inhibits activation of NF-κB signaling --- p.115 / Chapter 3.3 --- Discussion --- p.117 / Chapter CHAPTER FOUR --- THE PROTECTIVE ROLE OF MICRORNA-29B IN DIABETIC NEPHROPATHY --- p.121 / Chapter 4.1 --- Introduction --- p.122 / Chapter 4.2 --- Materials and methods --- p.123 / Chapter 4.2.1 --- Animal model --- p.123 / Chapter 4.2.2 --- Ultrasound-microbubble-mediated-miR-29 gene transfer --- p.124 / Chapter 4.2.3 --- Real-time polymerase chain reaction (PCR) --- p.124 / Chapter 4.2.4 --- Western Blotting --- p.125 / Chapter 4.2.5 --- Albumin excretion measurement --- p.126 / Chapter 4.2.6 --- ELISA --- p.126 / Chapter 4.2.7 --- Histology and immunohistochemistry --- p.126 / Chapter 4.2.8 --- In Situ hybridization --- p.127 / Chapter 4.2.9 --- Cell culture --- p.128 / Chapter 4.2.10 --- Statistical analysis --- p.129 / Chapter 4.3 --- Results --- p.129 / Chapter 4.3.1 --- Over-expression of miR-29b attenuates, but knockdown of miR-29b enhances fibrosis in vitro --- p.129 / Chapter 4.3.1.1 --- Over-expression of miR-29b attenuates fibrosis --- p.129 / Chapter 4.3.1.2 --- Knockdown of miR-29b enhances fibrosis --- p.132 / Chapter 4.3.2 --- Restoring miR-29b attenuates kidney injury in db/db mice --- p.134 / Chapter 4.3.3 --- Restoring miR-29b attenuates renal fibrosis in db/db mice --- p.139 / Chapter 4.3.3.1 --- Restoring miR-29b attenuates collagen IV in db/db mice --- p.139 / Chapter 4.3.3.2 --- Restoring miR-29b attenuates collagen I in db/db mice --- p.141 / Chapter 4.3.3.3 --- Restoring miR-29b attenuates fibronectin in db/db mice --- p.144 / Chapter 4.3.4 --- Restoring miR-29b inhibits renal fibrosis via TGF-β/Smad3 dependent pathway --- p.146 / Chapter 4.3.5 --- Restoring miR-29b inhibits th1 immune response in diabetic kidney --- p.148 / Chapter 4.3.6 --- Restoring miR-29b inhibits inflammation in diabetic kidney --- p.151 / Chapter 4.4 --- Discussion --- p.154 / Chapter 4.5 --- Conclusion --- p.161 / Chapter CHAPTER FIVE --- THE PATHOGENIC ROLE OF C-REACTIVE PROTEIN IN DIABETIC NEPHROPATHY --- p.162 / Chapter 5.1 --- Introduction --- p.163 / Chapter 5.2 --- Materials and methods --- p.164 / Chapter 5.2.1 --- Mouse model of STZ induced diabetes --- p.164 / Chapter 5.2.2 --- Measurement of blood glucose, urinary albumin excretion, and creatinine clearance --- p.165 / Chapter 5.2.3 --- Histology and immunohistochemistry --- p.165 / Chapter 5.2.4 --- Cell culture --- p.166 / Chapter 5.2.5 --- Real-time PCR --- p.166 / Chapter 5.2.6 --- Western blotting --- p.167 / Chapter 5.2.7 --- Statistical analyses --- p.168 / Chapter 5.3 --- Results --- p.168 / Chapter 5.3.1 --- Diabetic renal injury is exacerbated in CRP Tg mice --- p.168 / Chapter 5.3.2 --- Renal inflammation is exacerbated in diabetic CRP Tg mice --- p.172 / Chapter 5.3.2.1 --- F4/80+ macrophage infiltration is enhanced in diabetic CRP Tg mice --- p.172 / Chapter 5.3.2.2 --- CD3+ T cell infiltration is enhanced in diabetic CRP Tg mice --- p.173 / Chapter 5.3.2.3 --- TNF-α expression is enhanced in diabetic CRP Tg mice --- p.173 / Chapter 5.3.2.4 --- IL-1β expression is enhanced in diabetic CRP Tg mice --- p.174 / Chapter 5.3.3 --- Renal fibrosis is enhanced in diabetic CRP Tg mice --- p.175 / Chapter 5.3.3.1 --- Collagen I is enhanced in Diabetic CRP Tg mice --- p.175 / Chapter 5.3.3.2 --- Collagen IV is enhanced in diabetic CRP Tg mice --- p.176 / Chapter 5.3.4 --- Enhanced CRP signaling and activation of NF-κB and TGF-β/Smad3 signaling are key mechanism by which CRP promotes diabetic renal injury --- p.177 / Chapter 5.3.4.1 --- Enhanced CRP signaling via upregulation of CD32a expression --- p.177 / Chapter 5.3.4.2 --- enhanced activation of NF-κB signaling is key mechanism by which CRP promotes renal inflammation --- p.179 / Chapter 5.3.4.3 --- Enhanced activation of TGF-β/Smad3 signaling is key mechanism by which CRP promotes renal inflammation --- p.181 / Chapter 5.4 --- Discussion --- p.194 / Chapter 5.5 --- Conclusion --- p.197 / Chapter CHAPTER SIX --- SUMMARY AND CONCLUSION --- p.198 / REFERENCES --- p.202

2 型糖尿病(T2D)的發展歷史悠久，但導致T2D 患者胰島素抵抗的確切病理還沒有完全理解。骨骼肌佔大多數(70-80%)的胰島素引導的葡萄糖的吸收，所以它一直是胰島素抵抗的研究焦點。許多 T2D 患者的骨骼組織顯示線粒體功能障礙，但線粒體功能障礙和胰島素抵抗之間的關係尚不清楚還在辯論中。在這個項目中，這種關係是通過研究游離脂肪酸(FFA)( 24 小時處理)對 C2C12 小鼠骨骼肌細胞的效果來闡明。 / 免疫印記法顯示FFA 誘導胰島素抵抗，結合二維電泳和質譜分析的蛋白質組學研究發現FFA 有抑制糖酵解，增加β-氧化作用，沒有改變檸檬酸循環和抑製氧化磷酸化的作用。FFA 抑制電子傳遞鏈的幾個組成部分，揭示線粒體功能障礙，背後的原因可推測為FFA 增加令β-氧化作用增加，但沒有協調改變率檸檬酸循環，導致積累不完全β-氧化的中轉體，導致線粒體過載，最終導致胰島素抵抗。 / There is a long history of Type 2 diabetes (T2D) research development, but the exact pathology leading to insulin resistance of T2D is still not fully understood. T2D is frequently characterized by tissue insulin resistance and it is often associated with an elevated concentration of palmitic acid (PA, a major kind of dietary fatty acid) in serum. Due to this correlation, much of the effort in the field had been concentrated on the effect of PA in insulin action and glucose metabolism, and how elevated PA could possibly cause insulin resistance in specific tissues. / Skeletal muscle accounts for the majority (70-80%) of insulin-mediated glucose uptake, so it has been the focus of insulin resistance studies. Many T2D patients having elevated serum free fatty acid (FFA, where PA is a kind of FFA) also show mitochondrial dysfunction in their skeletal tissue, but the relationship between mitochondrial dysfunction and insulin resistance in skeletal muscle remains unclear and under debate. In this project, the three-party relationship was elucidated by studying the effect of 24hrs of incubation of palmitic acid (PA) on skeletal muscle using C2C12 mouse skeletal cells as model. / PA-treated C2C12 cells show reduction in insulin-stimulated Akt phosphorylation when compared with untreated C2C12 cells. Comparative proteomic study for both total proteins and mitochondrial proteins with 2D gel electrophoresis and mass spectrometry unveil, when compared with untreated cells, PA-treated C2C12 cells show down-regulation in enzymes involved in glycolysis(e.g. glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, fructose-bisphosphate aldose A), up-regulation in enzymes involved in beta-oxidation(e.g. 3-ketoacyl-CoA thiolase, 3-hydroxyacyl-CoA dehydrogenase), and down-regulation in proteins involved in oxidative phosphorylation(e.g. ATP synthase subunits, NADH-ubiquinone oxidoreductase 75kDa subunit, cytochrome b-c complex subunit 1). / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Lam, Chor Kwan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 69-78). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts also in Chinese. / Thesis/Assessment Committee --- p.i / Declaration --- p.ii / Abstract (in English) --- p.iii / Abstract (in Chinese) --- p.v / Acknowledgments --- p.vi / Table of Contents --- p.vii / List of Abbreviations --- p.x / List of Figures --- p.xiii / List of Tables --- p.xiv / Chapter 1. --- Literature review --- p.1 / Chapter 1.1. --- Introduction to diabetes mellitus --- p.1 / Chapter 1.1.1. --- Definition and prevalence --- p.1 / Chapter 1.1.2. --- Diagnosis and classification --- p.2 / Chapter 1.1.3. --- Symptoms and complications --- p.4 / Chapter 1.1.4. --- Causes and risk factors --- p.5 / Chapter 1.1.5. --- Prevention and treatment --- p.6 / Chapter 1.2. --- The role of muscle tissue in pathophysiology of T2DM --- p.7 / Chapter 1.3. --- Insulin receptor substrate-1 and Fatty acids-induced insulin resistance --- p.15 / Chapter 1.4. --- Introduction of proteomics --- p.18 / Chapter 1.4.1. --- The application of proteomics in disease discovery --- p.18 / Chapter 1.4.2. --- Application of Proteomics --- p.19 / Chapter 1.4.3 --- Two-dimensional gel electrophoresis --- p.20 / Chapter 1.4.4 --- Organelles proteomics --- p.21 / Chapter 1.4.5. --- Mass spectrometry --- p.22 / Chapter 1.4.6 --- Application of proteomic technology in disease pathology --- p.24 / Chapter 1.4.7 --- Current challenges --- p.25 / Chapter 1.5 --- Objectives --- p.27 / Chapter 2 --- Materials and Methods --- p.28 / Chapter 2.1 --- Fatty acid preparation --- p.28 / Chapter 2.2 --- Cell culture --- p.28 / Chapter 2.2.1 --- Treatment of C2C12 myotubes with Palmitic acid --- p.28 / Chapter 2.2.2 --- MTT assay for viability measurement --- p.29 / Chapter 2.2.3 --- Determination of the IC₅₀ values --- p.31 / Chapter 2.3 --- Proteomic analysis of C2C12 cells with and without PA treatment --- p.32 / Chapter 2.3.1 --- Protein sample preparation from C2C12 skeletal muscle cells --- p.32 / Chapter 2.3.2 --- Protein quantitation --- p.33 / Chapter 2.3.3 --- 2D Gel electrophoresis --- p.34 / Chapter 2.3.4 --- Image analysis --- p.36 / Chapter 2.3.5 --- In gel digestion and MALDI-ToF MS --- p.37 / Chapter 2.4 --- Mitochondrial purification and protein extraction --- p.38 / Chapter 2.4.1 --- Ultracentrifugation method --- p.38 / Chapter 2.4.2 --- Mitochondrial Isolation Kit --- p.39 / Chapter 2.5 --- Western Immunoblotting --- p.40 / Chapter 2.5.1 --- Protein sample preparation --- p.40 / Chapter 2.5.2 --- SDS-PAGE --- p.40 / Chapter 2.5.3 --- Western blotting --- p.40 / Chapter 2.5.4 --- Membrane Blocking and Antibody Incubations --- p.41 / Chapter 2.5.5 --- Detection of Proteins --- p.42 / Chapter 3 --- Results --- p.43 / Chapter 3.1 --- Differentiation of C2C12 myoblast into myotubes --- p.43 / Chapter 3.2 --- The effect of Palmitic acid on C2C12 Proliferation --- p.44 / Chapter 3.3 --- Comparison of total protein profiles of palmitic acid-treated C2C12 myotubes with control myotubes --- p.45 / Chapter 3.4 --- Western blotting of Akt and Phospho-Akt in C2C12 cells treated with Palmitic acid after acute exposure to insulin --- p.50 / Chapter 3.5 --- Comparison of two mitochondria isolation methodsultracentrifugation and mitochondrial isolation kit --- p.51 / Chapter 3.5.1 --- Quantity of extracted mitochondrial protein --- p.51 / Chapter 3.5.2 --- Purity of extracted mitochondrial protein --- p.52 / Chapter 3.6 --- Comparison of mitochondrial protein profiles between palmitic acid-treated and control C2C12 myotubes --- p.53 / Chapter 3.7 --- Western blotting of insulin receptor substrate-1 and its serine phosphorylation --- p.58 / Chapter 4 --- Discussion --- p.59 / Chapter 4.1 --- Investigation of anti-proliferating effect of Palmitic acid on C2C12 using MTT assay --- p.59 / Chapter 4.2 --- Comparison of total protein profiles of palmitic C2C12 myotubes with control myotubes --- p.60 / Chapter 4.3 --- Western blotting of insulin receptor substrate-1and its serine phosphorylation --- p.62 / Chapter 4.4 --- Western blotting of Akt and Phospho-Akt in C2C12 cells treated with Palmitic acid after acute exposure to insulin --- p.63 / Chapter 4.5 --- Comparison of mitochondrial protein profiles between palmitic acid-treated and control C2C12 myotubes --- p.65 / Chapter 4.6 --- Problems faced and future prospect --- p.68 / Chapter 5 --- References --- p.69

Introdução: Não foram encontrados na literatura dados sobre a prevalência de diabetes mellitus em mães de crianças com fissura labiopalatina. Dada a relevância do tema esse foi o principal objetivo da presente investigação. Método: Após aprovação do comitê de ética e pesquisa e obtenção do consentimento livre e esclarecido, foram entrevistadas 325 mulheres, mães biológicas de crianças com fissuras labiopalatinas com idades de 0 a 3 anos matriculadas no Hospital de Reabilitação de Anomalias Craniofaciais da Universidade de São Paulo, HRAC-USP. Foi aplicado questionário com 24 questões onde obteve-se a identificação das mães, o tipo de diabetes, os principais sinais clínicos, as comorbidades associadas, hipertensão, obesidade, medicamentos e drogas lícitas e ilícitas usadas durante a gravidez. Também foram aferidos a pressão arterial e o perímetro abdominal das mães e anotados os valores da glicemia de jejum na primeira consulta pré-natal. Resultados: Os resultados obtidos mostraram que 88 mulheres apresentavam diabetes mellitus, sendo 78 com diabetes mellitus gestacional, 05 com diabetes mellitus tipo 1 e 05 com diabetes mellitus tipo 2. A prevalência de diabetes mellitus em mães de crianças com fissuras labiopalatinas foi de 27,08%, comparativamente maior (p<0,01) aos valores encontrados na população adulta brasileira (7,6%). Dessas 88 mulheres, foram excluídos os fatores que de alguma forma pudessem influenciar o aparecimento de fissuras labiopalatinas e obteve-se o percentual de 16%, onde a hiperglicemia materna nessas mães foi o único fator provável no aparecimento dessas fissuras. Conclusão: A prevalência de diabetes mellitus em mães de crianças com fissuras labiopalatinas foi de 27,08%. (p<0,01). Extraídos os fatores tidos como confundidores, tais como medicamentos usados na gestação, álcool, tabaco, drogas ilícitas, obesidade e hipertensão arterial, restaram 52 mães que tinham somente a hiperglicemia materna como fator isolado, num total de 16%. / Introduction: there was no data found on the literature regarding the prevalence of diabetes mellitus in mothers of children born with cleft lip and palate. Given the relevance of the subject, that was the main objective of this investigation. Method: after approval by the research and ethics committee, and the free, informed consent firmed by the mothers, interviews were conducted with 325 women biological mothers of children aged between 0 to 3 years, born with cleft lip and palate, registered at Hospital for Rehabilitation of Craniofacial Anomalies, University of São Paulo, HRAC-USP. A 24-question survey was applied to collect data regarding the mothers identification, type of diabetes, clinical data, associated comorbidities, hypertension, obesity, medication and legal or illegal drugs used during pregnancy. Blood pressure and abdominal circumference were measured, and fasting blood glucose levels measured at the first prenatal visit. Results: It was found that 88 women had diabetes mellitus, 78 of which had gestational diabetes, 05 had type 1 diabetes and 05 had type 2 diabetes the prevalence of diabetes in mothers of children with cleft lip and palate was 27.08%, a number significantly higher (p<0.01) than the rates found among the Brazilian adult population (7.6%). From this group of 88 women we have suppressed the factors that could contribute to the occurrence of cleft lip and palate in some way, obtaining a prevalence of 16% in which maternal hyperglycemia was the only likely cause of cleft lip and palate. Conclusion: The prevalence of diabetes mellitus in mothers of children born with cleft lip and palate was 27.08% (p<0.01). Suppressing other potentially confusing factors like medication used during pregnancy, alcohol, tobacco, illegal drugs, obesity and hypertension, there were 52 mothers who had only maternal hyperglycemia as an isolated factor, representing 16% of the total studied population.

Lee, Man Yuen. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 191-215). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Master of Public Health / OBJECTIVES: In Australia, diabetes causes significant morbidity and mortality. Whilst the need to prevent diabetes and its complications has been widely recognised, the capacity of health care systems - which organise diabetes care - to facilitate prevention has not been fully established. METHODS: A series of seven population-based case-control studies were used to examine the effectiveness of the Australian health care system and its capacity to manage diabetes. Six of the studies compared the patterns of care of patients who had developed advanced diabetes complications in 2000 (cases), to similar patients who remained free of the condition (controls) across Australia and for various risk groups. A secondary study investigated the role of treating GPs in the development of the outcome. RESULTS: A strong relationship between the patterns of care and the development of advanced diabetes complications was found and is described in Chapter 4. In Chapter 5, this same relationship was investigated for each Australian state and territory, and similar findings were made. The study in Chapter 6 investigated whether late diagnosis or the patterns of care was the stronger risk factor for advanced diabetes complications, finding that the greatest risk was associated with the latter. In Chapter 7 the influence of medical care during the pre-diagnosis period was explored, and a strong relationship between care obtained in this period and the development of advanced complications was found. In Chapter 8, which investigated the role of socio-economic status in the development of advanced complications, found that the risk of advanced diabetes complications was higher in low socio-economic groups. Chapter 9 investigated geographic isolation and the development of advanced diabetes complications and found that the risk of advanced complications was higher in geographically isolated populations. Finally, Chapter 10, which utilised a provider database, found that some GP characteristics were associated with the development of advanced diabetes complications in patients. CONCLUSION: A number of major risk factors for the development of advanced complications in Australia was found. These related to poorer diabetes management, later diagnosis, low socioeconomic status and geographic isolation. Strategies must be devised to promote effective diabetes management and the early diagnosis of diabetes across the Australian population.

People with diabetes are at a greater risk of burn injuries than those without diabetes. This stems from the epidemiological profiles of the conditions and the effects of morbidities associated with diabetes. Both conditions share some similarities in terms of metabolic alterations and suboptimal immune functions which may result in poor outcomes for patients. For that reason, it is reasonable to deduce that patients with diabetes are a challenging group to manage in burns units. However, this deduction should be taken cautiously because of lack of supporting evidence. Accordingly and after consulting with clinical experts, the research in this portfolio investigated the association between diabetes and burn injuries. In particular, two different aspects of this association were investigated in two individual quantitative and descriptive inquiries. The first was a case note review of patients hospitalised with a principal diagnosis of a foot burn injury in a large tertiary hospital in South Australia from 1999 to 2004. The second study investigated management of diabetes in burns units treating adults. This study is an e-mail survey of clinical leaders of burns units in Australia, New Zealand, Hong Kong and the United Kingdom. The clinical leaders were approached indirectly through key liaison persons in each identified unit. In the first study, outcomes for twelve subjects with and fifty-two without diabetes were described using descriptive and non-parametric statistics. In the second study, descriptive frequencies and content analysis were adopted to analyse twenty-nine responses from seventeen out of thirty burns units which participated in the study. Supporting findings in the literature, the first study showed that burn injuries among subjects with diabetes were mainly resulted from household devices. There were no statistically significant differences between subjects with and without diabetes in terms of size and depth of burn injuries and treatment received. In spite of this, there was a statistically significant association between diabetes and the experience of local post-burn complications and longer duration of hospitalisation. The second study indicated that more than twenty-five percent of the respondents believed that multidisciplinary centres should only occasionally be involved in the process of care. Participants reported that the individual profile of each patient plays a major role in determining the management of diabetes. Additionally, it was found that the insulin sliding scale was commonly used in the management of diabetes in burns units. The association between diabetes and a burn injury is a serious issue in terms of health and cost. This association need be addressed firstly and most importantly at the prevention level; secondly through proper management of both diabetes and burns. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1285462 / Thesis (D.Nurs.)--Population Health and Clinical Practice, 2007.

Type 2 diabetes mellitus (T2DM) is a disease of multi-complications affecting more than 20 million US adults. Hyperglycemia is the classic clinical feature of diabetes, and uncontrolled hyperglycemia leads to deadly health complications. Thus, control of blood glucose represents a major goal for diabetics. Human and rodent studies revealed another clinical feature; diabetics have low tissue and plasma levels of polyunsaturated fatty acids (PUFAs), an effect often attributed by impaired endogenous PUFA synthesis. In this context, rodent studies have revealed a possible link between PUFA synthesis and high fat diet induced obesity and diabetes. These studies have shown that obese and diabetic mice have low hepatic expression and activity of fatty acid elongase-5 (Elovl5), a key enzyme involved in the PUFA synthesis pathway. Over-expression of Elovl5 in livers of chow fed C57BL/6J mice decreased fasting blood glucose and increased hepatic glycogen contents. Therefore, my hypothesis for the current work is that elevated hepatic Elovl5 activity or improved hepatic PUFA synthesis will improve systemic and hepatic carbohydrate metabolism in a mouse model of diet induced obesity and diabetes. Using a recombinant adenovirus approach, we over-expressed Elovl5 in livers of high fat diets (60% calories derived from fat as lard, Research Diets) induced obese-diabetic mice. Elevated hepatic Elovl5 activity increased hepatic and plasma C��������������� PUFA contents, reduced homeostatic model assessment for insulin resistance (HOMA-IR), improved glucose tolerance and lowered fasting blood glucose to euglycemic levels in obese-diabetic mice. The mechanism for insulin mimetic effect of Elovl5 on hepatic glucose metabolism was correlated with increased phosphorylation of Akt-S��������, FoxO1-S�������� and PP2Acat-Y��������, decreased nuclear content of FoxO1, and decreased expression of Pck1 and G6Pase; important enzymes involved in gluconeogenesis (GNG) and glucose production. Phospho-FoxO1 is excluded from nuclei, ubiquitinated and degraded by the proteasome. Loss of nuclear FoxO1, due to its increased phosphorylation, leads to the reduction in the expression of key genes involved in gluconeogenesis, i.e., Pck1 and G6Pase. Using obese-diabetic mice liver extracts and HepG2 cells, I established that Elovl5 uses two mechanisms to control hepatic GNG. The first mechanism involves Elovl5 mediated increased Akt2-S�������� and FoxO1-S�������� phosphorylation via mTORC2-rictor pathway. The second mechanism involves Elovl5 mediated attenuation of de-phosphorylation of FoxO1 via PP2A inhibition. Together, these mechanisms increase FoxO1 phosphorylation status in livers of fasted obese-diabetic mice, lower hepatic FoxO1 nuclear abundance and FoxO1 capacity to sustain transcription of GNG genes and inhibit GNG and restore blood glucose levels in fasted obese-diabetic mice. Results of these studies showed Elovl5 corrected high fat diet induced hyperglycemia in C57BL/6J mice, identified the molecular mechanism of Elovl5 control of GNG and explained how Elovl5 or PUFA synthesis controls GNG. Therefore, these findings will be eventually helpful in developing a therapeutic target to combat hyperglycemia. / Graduation date: 2013 / Access restricted to the OSU Community at author's request from July 27, 2012 - July 27, 2013

published_or_final_version / Nursing Studies / Master / Master of Nursing in Advanced Practice