Postprandial urea production in subjects with insulin dependent diabetes mellitus (IDDM) on conventional insulin therapy is normal when the previous diet is high in protein, but there is an incomplete adaptive reduction in urea production following protein restriction. To evaluate the nutritional implications of restricted protein intake in human diabetes mellitus, it is first necessary to establish a reliable method to measure changes in urea production and amino acid catabolism in response to changes in dietary protein intake. We therefore tested (1) the accuracy of the urea production rate (Ra) to depict changes in urea production, (2) whether sulfate production can be accurately depicted using tracer or nontracer approaches, after establishing the use of electrospray tandem mass spectrometry to measure sulfate concentrations and 34SO4 enrichments following administration of the stable isotope tracer sodium [34S]sulfate, (3) the reproducibility of urea and sulfate measurements following a test meal low in protein (0.25 g/kg) in subjects previously adapted to high (1.5 g/kg.d) and low (0.3 g/kg.d) protein intakes, and compared the metabolic fate of [ 15N]alanine added to the test meal with that of [15N] Spirulina platensis, a 15N-labeled intact protein, and (4) whether we could identify the differences in postprandial urea and sulfate productions between normal subjects and persons with IDDM receiving conventional insulin therapy previously adapted to high protein intake, when the test meal was limiting in protein. Under basal conditions, steady state urea Ra is an accurate measure of urea production. Following changes in urea production, both the tracer and nontracer methods seriously underestimated total urea Ra. The tracer method overestimated sulfate production by 20%, but the nontracer method provided an accurate measure of sulfate production and, hence, sulfur amino acid catabolism. Postprandial changes in urea and sulfate productions following normal ada

Glutamate formiminotransferase deficiency (OMIM 229100) is an autosomal recessive disorder marked by clinical heterogeneity. The severe phenotype, first identified in patients of Japanese descent, includes high levels of formiminoglutamate (FIGLU) in the urine in response to histidine loading, megaloblastic anemia, and mental retardation. The mild phenotype is marked by high levels of FIGLU in the urine in the absence of histidine loading, mild developmental delay and no hematological abnormalities. The gene for human glutamate formiminotransferase-cyclodeaminase consists of 15 exons and is located at 21q22.3. The protein consists of a tetramer of dimers, with dimerization essential for both formiminotransferase and cyclodeaminase activity. / Genomic DNA extracted from cell lines from three patients with suspected glutamate formiminotransferase deficiency was analyzed by PCR and sequencing of individual exons. Cell lines WG 1758 and WG 1759 are from two siblings of Germanic descent. Both siblings are heterozygous for the mutations c457 C → T and c940 G → C. The c457 C → T changes a conserved arginine to a cysteine in a loop involved in the binding of formiminotetrahydrofolate to the enzyme. The c940 G → C mutation converts an arginine to a proline in an alpha-helix essential for the dimerization of the formiminotransferase domain. Cell line WG 1795 is from a patient of Danish descent. The patient appears to be hemizygous for a c1033 insG mutation. Quantitative PCR suggests the presence of a deletion on the other chromosome, which minimally encompasses exon 9. All of the FTCD gene changes were absent in 100 control individuals (200 alleles).

Thesis (Ph. D.)--University of Washington, 2004. / Vita. Includes bibliographical references (leaves 152-161).

Made available in DSpace on 2014-08-13T14:50:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-22Bitstream added on 2014-08-13T18:00:39Z : No. of bitstreams: 1 000756842.pdf: 789344 bytes, checksum: 061035505a8bd708cd9dbac41aad0d39 (MD5) / A obesidade, doença crônica metabólica caracterizada pelo acúmulo excessivo de tecido adiposo em relação à massa magra tecidual, é considerada uma epidemia global e um importante problema de saúde pública, que afeta tanto países desenvolvidos quanto subdesenvolvidos. O adipócito recebe influência de diversas substâncias e secreta inúmeros peptídeos, como leptina, angiotensina I e II, TGF-β, entre outros, que atuam diretamente ou indiretamente no sistema cardiovascular. Assim, o tecido adiposo não é simplesmente um reservatório de energia, mas um ativo órgão endócrino, parácrino e autócrino com múltiplas funções, capaz de sintetizar e liberar mediadores que participam de diversos processos biológicos, incluindo os que ocorrem no coração. O coração é composto por miócitos, nervos, vasos e matriz extracelular. O principal componente da matriz é o colágeno, com predomínio dos tipos I e III, sendo que, o tipo I é o mais abundante, correspondendo á aproximadamente 80% do colágeno total miocárdico. O colágeno é produzido, principalmente, pelos fibroblastos e degradado pelas metaloproteinases (MMPs). O colágeno, em situação estável, contribui para a manutenção da arquitetura e função cardíaca; entretanto, em resposta a estímulos desencadeados por agentes neuro-hormonais e/ou mecânicos, pode sofrer alterações; esta mudança pode ser resultante do aumento da síntese e/ou diminuição da degradação ou vice-versa. Em pesquisa recente realizada em nosso laboratório foi encontrado diminuição dos níveis protéicos de colágeno tipo I miocárdico em ratos Wistar obesos, por dieta hiperlipídica insaturada por 30 semanas, em relação ao grupo controle. Em razão, dos resultados encontrados em nosso laboratório e da literatura mostrar que a leptina aumenta a atividade das MMP-2 e a síntese da expressão gênica da MMP-9, a proposta deste estudo foi testar a hipótese ... / Obesity is a chronic metabolic disorder characterized by excessive adipose tissue accumulation in relation to lean tissue. Currently, it is a global epidemic and a major public health problem that affects both developed as well as undeveloped countries. The adipocyte receives influence of several substances and secretes numerous peptides, such as leptin, angiotensin I and II, TGF-β, among others, that act directly or indirectly on the cardiovascular system. Thus, adipose tissue is not simply an energy reservoir, but an active endocrine, paracrine and autocrine organ with multiple functions, able to synthesize and release mediators that participate in many biological processes, including those that occur in the heart. The heart is composed of myocytes, nerves, vessels and extracellular matrix. The main component of matrix is collagen, predominantly type I and III, being type I the most abundant, corresponding to approximately 80% of total myocardial collagen. Collagen is mainly produced by fibroblasts and degraded by metalloproteinases (MMPs). Collagen, in a stable condition, contributes to the maintenance of cardiac architecture and function, however, in response to stimuli triggered by neuro-hormonal and/or mechanical agents, it may change, and this change can be due to increased synthesis and/or decreased degradation, or vice versa. In recent research conducted in our laboratory, we found decreased protein levels of myocardial type I collagen in obese Wistar rats by unsaturated high-fat diet for 30 weeks. Due to the results found in our laboratory and because the literature shows that leptin increases MMP-2 activity and MMP-9 gene expression, the purpose of this study was to test the hypothesis that the reduction of myocardial type I collagen is associated with increased MMPs 2 and 9 activities in obese rats by unsaturated high-fat diet. Thirty-day-old male Wistar rats were randomized into to two groups: control ...

Made available in DSpace on 2014-06-11T19:32:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-01-13Bitstream added on 2014-06-13T20:27:54Z : No. of bitstreams: 1 siloto_ev_dr_botfmvz.pdf: 494737 bytes, checksum: 2225a359b9e80a74e8528c9b0ea4cfd8 (MD5) / O presente estudo avaliou o efeito da suplementação do cromo e dois níveis de energia sobre o desempenho e a qualidade de ovos de poedeiras leves. Foram utilizadas 192 poedeiras da linhagem Bovans com 47 semanas de idade. As aves foram distribuídas em um delineamento inteiramente casualizado, em esquema fatorial 4x2, com quatro níveis de suplementação de cromo (0, 200, 400 e 800 μg Cr/kg) e dois níveis energéticos (2.780 e 2.900 kcal EM/kg de ração). Cada tratamento continha seis repetições de quatro aves cada. O cromo utilizado foi o derivado de levedura. O experimento teve a duração de 112 dias, divididos em 4 períodos de 28 dias. Os parâmetros de desempenho não foram influenciados pelos níveis de cromo e de energia da dieta. Houve aumento linear no percentual de gema com o aumento da suplementação de cromo em dietas contendo 2.780 kcal/kg. As aves alimentadas com dietas contendo 2.780 kcal de EM/kg e isentas de suplementação de cromo, apresentaram redução no percentual de gema. Quando as aves foram suplementadas com 800 μg Cr/kg de dieta, foi observada uma maior deposição de gema. A suplementação de 800 μg Cr/kg na dieta com 2.780 kcal EM/kg provocou aumento no percentual de gema sem afetar o desempenho das poedeiras / This study aimed to evaluate the performance and egg quality in laying hens receiving diets supplemented with crescent levels of Chromium yeast and two different dietary energy levels. One hundred and ninety-two 47-week-old Bovans laying hens were distributed in a completely randomized design with 4x2 factorial arrangement: 4 levels of chromium yeast supplementation (0, 200, 400, 800 μg/kg feed) and two energy levels (2,780 and 2,900 kcal ME / kg of feed). Each treatment consisted of 6 replications of 4 birds each. The experiment lasted 112 days (four 28-day periods). The following performance parameters were evaluated: feed intake, egg weight, egg production, percentage of whole eggs, egg mass, feed: gain ratio (kg feed/dozen eggs and kg feed/kg eggs). The egg quality was accessed through the weight of total egg, shell, yolk and albumen and their proportions, Haugh unit, the specific gravity, and the shell thickness. The performance parameters were not influenced by the levels of chromium and dietary energy. There was a linear increase in the percentage of yolk with increasing chromium supplementation in diets containing 2.780kcal/kg. Laying hens fed with diets containing 2.780kcal/kg ME and without chromium supplementation showed reduced percentage of yolk. When birds were supplemented with 800 mg Cr / kg diet a greater deposition of yolk was observed. Dietary supplementation of 800 mg Cr / kg with 2.780kcal of ME/kg induced an increase in the percentage of yolk without affecting performance of laying hens

Orientador: Antonio Carlos Cicogna / Banca: Marcos Ferreira Minicucci / Banca: André Soares Leopoldo / Resumo: A obesidade, doença crônica metabólica caracterizada pelo acúmulo excessivo de tecido adiposo em relação à massa magra tecidual, é considerada uma epidemia global e um importante problema de saúde pública, que afeta tanto países desenvolvidos quanto subdesenvolvidos. O adipócito recebe influência de diversas substâncias e secreta inúmeros peptídeos, como leptina, angiotensina I e II, TGF-β, entre outros, que atuam diretamente ou indiretamente no sistema cardiovascular. Assim, o tecido adiposo não é simplesmente um reservatório de energia, mas um ativo órgão endócrino, parácrino e autócrino com múltiplas funções, capaz de sintetizar e liberar mediadores que participam de diversos processos biológicos, incluindo os que ocorrem no coração. O coração é composto por miócitos, nervos, vasos e matriz extracelular. O principal componente da matriz é o colágeno, com predomínio dos tipos I e III, sendo que, o tipo I é o mais abundante, correspondendo á aproximadamente 80% do colágeno total miocárdico. O colágeno é produzido, principalmente, pelos fibroblastos e degradado pelas metaloproteinases (MMPs). O colágeno, em situação estável, contribui para a manutenção da arquitetura e função cardíaca; entretanto, em resposta a estímulos desencadeados por agentes neuro-hormonais e/ou mecânicos, pode sofrer alterações; esta mudança pode ser resultante do aumento da síntese e/ou diminuição da degradação ou vice-versa. Em pesquisa recente realizada em nosso laboratório foi encontrado diminuição dos níveis protéicos de colágeno tipo I miocárdico em ratos Wistar obesos, por dieta hiperlipídica insaturada por 30 semanas, em relação ao grupo controle. Em razão, dos resultados encontrados em nosso laboratório e da literatura mostrar que a leptina aumenta a atividade das MMP-2 e a síntese da expressão gênica da MMP-9, a proposta deste estudo foi testar a hipótese ... / Abstract: Obesity is a chronic metabolic disorder characterized by excessive adipose tissue accumulation in relation to lean tissue. Currently, it is a global epidemic and a major public health problem that affects both developed as well as undeveloped countries. The adipocyte receives influence of several substances and secretes numerous peptides, such as leptin, angiotensin I and II, TGF-β, among others, that act directly or indirectly on the cardiovascular system. Thus, adipose tissue is not simply an energy reservoir, but an active endocrine, paracrine and autocrine organ with multiple functions, able to synthesize and release mediators that participate in many biological processes, including those that occur in the heart. The heart is composed of myocytes, nerves, vessels and extracellular matrix. The main component of matrix is collagen, predominantly type I and III, being type I the most abundant, corresponding to approximately 80% of total myocardial collagen. Collagen is mainly produced by fibroblasts and degraded by metalloproteinases (MMPs). Collagen, in a stable condition, contributes to the maintenance of cardiac architecture and function, however, in response to stimuli triggered by neuro-hormonal and/or mechanical agents, it may change, and this change can be due to increased synthesis and/or decreased degradation, or vice versa. In recent research conducted in our laboratory, we found decreased protein levels of myocardial type I collagen in obese Wistar rats by unsaturated high-fat diet for 30 weeks. Due to the results found in our laboratory and because the literature shows that leptin increases MMP-2 activity and MMP-9 gene expression, the purpose of this study was to test the hypothesis that the reduction of myocardial type I collagen is associated with increased MMPs 2 and 9 activities in obese rats by unsaturated high-fat diet. Thirty-day-old male Wistar rats were randomized into to two groups: control ... / Mestre

Orientador: Eduardo Rochete Ropelle / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Aplicadas / Made available in DSpace on 2018-08-24T14:06:26Z (GMT). No. of bitstreams: 1 Pauli_LucianaSantosSouza_M.pdf: 2136192 bytes, checksum: ba071049e1cda0b7eb5230076d5d4b4e (MD5) Previous issue date: 2013 / Resumo: A proteína MKP-3 (MAPK phosphatase-3) possui capacidade de associar-se fisicamente e desfosforilar a FoxO1 no fígado, resultando em aumento da gliconeogênese e hiperglicemia na condição de obesidade e diabetes. Por outro lado, o exercício físico é uma das estratégias não farmacológicas mais utilizadas para a melhora do perfil glicêmico em pacientes diabéticos, no entanto, suas ações no tecido hepático são pouco conhecidas. Assim, o objetivo deste trabalho foi investigar os efeitos do treinamento físico sobre a expressão proteica da MKP-3, bem como sua interação com a FoxO1 em fígado de animais obesos. Ademais, tem como objetivo avaliar o mecanismo pelo qual o exercício físico é capaz de inibir a proteína MKP-3 em tecido hepático. Foram utilizados camundongos Swiss machos que receberam por 16 semanas uma dieta padrão ou rica em gordura. Os animais foram distribuídos aleatoriamente em seis grupos: controle (C), obeso sedentário (OB), obeso exercitado (EXE), obeso tratado com oligonucleotídeo antisense MKP-3 (OB-ASO), obeso tratado com sense MKP-3 (OB-sense) e obeso submetido ao treinamento físico e ao tratamento com antisense MKP-3 concomitantemente (OB-EXE-ASO). O protocolo de treinamento físico consistiu de natação com sessões de 60 minutos, 5 vezes por semana, com carga equivalente a 5% da massa corporal total do animal. O tratamento com antisense foi realizado através de injeções duas vezes ao dia, com um volume total de 2.0 ?l por dose (4.0 nmol/?l) por 5 dias. Ao final do experimento foram realizados os testes de tolerância ao piruvato (TTP) e teste de tolerância à insulina (TTI). Além disso, foram avaliados parâmetros fisiológicos (ingestão calórica, massa corporal, glicemia e insulinemia de jejum) e a análise molecular das proteínas (MKP-3, FoxO1, PGC1?, HNF-4?, PEPCK, G6Pase, ERK e CK2?) através das técnicas de imunoblot e imunoprecipitação. Utilizou-se a técnica de imunohistoquímica para análise da co-localização da MKP-3 no fígado dos animais. Os resultados obtidos demonstraram que o treinamento físico diminuiu expressão proteica de MKP-3 e a associação FoxO1/MKP-3 no fígado dos animais obesos. Além disso, camundongos obesos treinados apresentaram maiores níveis de FoxO1 fosforilada e diminuição da expressão das proteínas PGC-1? e HNF-4???no fígado se comparado aos animais obesos não submetidos ao treinamento físico. Verificam-se também menores níveis das enzimas gliconeogênicas PEPCK e G6Pase nos animais obesos treinados se comparado aos seus pares sedentários. Estes resultados a nível molecular foram acompanhados por alterações fisiológicas incluindo aumento da sensibilidade à insulina no fígado, menor produção hepática de glicose e redução da hiperglicemia nos camundongos obesos, independentemente da redução da massa corporal total. Ademais, não foram encontrados efeitos aditivos sobre os parâmetros fisiológicos e moleculares com o tratamento de oligonucleotídeo antisense MKP-3 e treinamento físico realizado concomitantemente nos animais obesos. Por fim, os efeitos supressivos do treinamento físico sobre a proteína MKP-3 parecem estar relacionados, no mínimo em parte, a diminuição na fosforilação das ERKs no fígado de camundongos obesos / Abstract: The protein MKP-3 (MAPK phosphatase-3) has the ability to physically associate with and dephosphorylate the FoxO1 in the liver, resulting in increased gluconeogenesis and hyperglycemia in the condition of obesity and diabetes. On the other hand, exercise is one of the most used non-pharmacological strategies for improving glycemic control in diabetic patients; however, their actions in liver tissue are poorly explored. The objective of this study was to investigate the effects of exercise on the expression of MKP-3 and its interaction with FoxO1 in the liver of obese animals. Furthermore, aimed at evaluating the mechanism by which physical exercise is able to inhibit protein MKP-3 in the liver. We used male Swiss mice that received a standard diet or high fat for 16 weeks. The animals were randomly divided into six groups: control (C), sedentary obese (OB), obese exercised (EXE), obese treated with MKP-3 antisense oligonucleotide (ASO-OB), obese treated with sense MKP-3 (OB- sense) and obese subjected to physical training and treatment with antisense MKP-3 concomitantly (OB-EXE-ASO). The protocol consisted of swimming exercise training with sessions of 60 minutes, five times per week with a load equivalent to 5% of the total body mass of the animal. Treatment with antisense was performed by injection twice daily, with a total volume of 2.0 ul per dose (4.0 nmol / microl) for 5 days. At the end of the experiment were performed to pyruvate tolerance tests (PTT) and insulin tolerance test (ITT). Furthermore, physiological parameters were evaluated (caloric intake, body weight, blood glucose and fasting insulin) and molecular analysis of proteins (MKP-3, FoxO1, PGC1a, HNF-4a PEPECK, G6Pase, ERK and CK2?) by immunoblotting and immunoprecipitation techniques. The immunohistochemistry technique was utilized to evaluate the co-localization of MKP-3 in the liver of the animals. The results showed that physical training decreased the protein expression of MKP-3 and association FoxO1/MKP-3. Further, trained obese mice had higher levels of phosphorylated FoxO1 and decreased PGC-1? and HNF-4??? There was also a decrease in the protein levels of PEPCK and G6Pase in the liver of mice exercised compared to sedentary obese mice. These results at molecular level were accompanied by physiological changes including increased insulin sensitivity in the liver, reduced hepatic glucose production and reduce hyperglycemia in obese mice, regardless of reducing total body mass. In addition, there were no additive effects of treatment with antisense oligonucleotide MKP-3 and physical training performed concurrently in obese animals. Finally, the suppressive effects of physical training on MKP-3 protein appear to be related at least in part, the decrease in phosphorylation of ERKs in the livers of obese mice / Mestrado / Metabolismo e Biologia Molecular / Mestra em Ciências da Nutrição e do Esporte e Metabolismo

No description available.

Ample studies have shown that perturbation of metabolic phenotype is correlated with gene mutation and pathogenesis of colorectal cancer (CRC) and rheumatoid arthritis (RA). Mass spectrometry (MS)-based metabolomics as a powerful and stable approach is widely applied to bridge the gap from genotype/metabolites to phenotype. In CRC suffers, KRAS mutation accounts for 35%-45%. In previous study, SLC25A22 that encodes the mitochondrial glutamate transporter was found to be overexpressed in CRC tumor and thus to be essential for the proliferation of CRC cells harboring KRAS mutations. However, the role of SLC25A22 on metabolic regulation in KRAS-mutant CRC cells has not been comprehensively characterized. We performed non-targeted metabolomics, targeted metabolomics and isotope kinetic analysis of KRAS-mutant DLD1 cells with or without SLC25A22 knockdown using ultra-high performance liquid chromatography (UHPLC) coupled to Orbitrap MS and tandem MS (MS/MS). In global metabolomics analysis, 35 differentially regulated metabolites were identified, which were primarily involved in alanine, aspartate and glutamate metabolism, urea cycle and polyamine metabolism. Then targeted metabolomics analysis on intracellular metabolites, including tricarboxylic acid (TCA) cycle intermediates, amino acids and polyamines, was established by using LC-MS/MS coupled with an Amide BEH column. Targeted metabolomics analysis revealed that most TCA cycle intermediates, aspartate (Asp)-derived asparagine, alanine and ornithine (Orn)-derived polyamines were strongly down-regulated in SLC25A22 knockdown cells. Moreover, the targeted kinetic isotope analysis using [U-13C5]-glutamine as isotope tracer showed that most of the 13C-labeled TCA cycle intermediates were down-regulated in SLC25A22-silencing cells. Orn-derived polyamines were significantly decreased in SLC25A22 knockdown cells and culture medium. Meanwhile, accumulation of Asp in knockdown of GOT1 cells indicated that oxaloacetate (OAA) was majorly converted from Asp through GOT1. Exogenous addition of polyamines could significantly promote cell proliferation in DLD1 cells, highlighting their potential role as oncogenic metabolites that function downstream of SLC25A22-mediated glutamine metabolism. SLC25A22 acts as an essential metabolic regulator during CRC progression as promotes the synthesis of TCA cycle intermediates, Asp-derived amino acids and polyamines in KRAS-mutant CRC cells. Moreover, OAA and polyamine could promote KRAS-mutant CRC cell growth and survival. Rheumatoid arthritis (RA) is a chronic, inflammatory and symmetric autoimmune disease and a major cause of disability. However, there is insufficient pathological evidence in term of metabolic signatures of rheumatoid arthritis, especially the metabolic perturbation associated with gut microbiota (GM). Based on consistent criteria without special diet and therapeutic intervention to GM, we enrolled 50 RA patients and 50 healthy controls. On basis of the platform of UHPLC-MS and GC-MS, were performed for the non-targeted metabolomics to investigate alterations of endogenous metabolites in response to RA inflammation and interaction with GM. 32 and 34 significantly changed metabolites were identified in urine and serum of patients with RA, respectively. The altered metabolites were identified by HMDB, METLIN database or authentic standards, and mostly metabolites were attributed into tryptophan and phenylalanine metabolism, valine, leucine and isoleucine biosynthesis, aminoacyl-tRNA biosynthesis and citrate cycle. To obtain alterations of more components in tryptophan and phenylalanine metabolism, we developed and validated a targeted metabolomics method of 19 metabolites by using LC-QqQ MS. Combining the results of targeted metabolomics with global metabolomics, significantly up-regulated kynurenine (KYN), anthranilic acid (AA) and 5-hydroxylindoleacetic acid (HIAA) simultaneously in urine and serum was found to implicate the activation of tryptophan metabolism under the condition of RA, which acted pro-inflammatory roles in inflammation and was closely correlated with GM. IDO/TDO functioned as a pro-inflammation mediator was overexpressed in RA patients. Urinary kynurenic acid and serum serotonin that have impacts on anti-inflammation in immune system were down-regulated in RA patients. The levels of phenylacetic acid and phenyllactic acid serving as a pro-inflammatory and an anti-inflammatory agent, respectively, increased in serum of patients with RA. Moreover, certain essential amino acids (EAAs), and mostly conditional EAAs were decreased in RA patients, which have been reported to inhibit cell proliferation of immune cells. In particular, deficiency of branched chain amino acids (BCAAs, valine and isoleucine) was observed in serum of patients with RA, which may lead to muscle loss and cartilage damage. The specificity of all altered metabolites resulted from RA was considerably contributed through the GM-derived metabolites. The findings revealed that GM-modulated RA inflammation was mainly resulted from tryptophan and phenylalanine metabolism, and amino acid biosynthesis, which may provide more information for better understanding the RA mechanism.

Indiana University-Purdue University Indianapolis (IUPUI) / Many cells are involved in the orchestra that is bone homeostasis--particularly osteoclasts and osteoblasts, which mediate remodeling of bones. This creates a balance that must be kept in check, otherwise pathologies arise. The JAK-STAT signaling pathway is crucial to maintaining this balance. It has long been known that the transcription factor STAT3 has more profound effects on bone homeostasis than other members of the STAT family of proteins. Recently, a genetic condition called Job’s Syndrome has been specifically linked to point mutations in the Stat3 gene. These patients present with severe bone abnormalities, including prominent foreheads, broad nasal bridges, and abnormal eye spacing. For this reason, our lab has extensively studied conditional knockouts of Stat3 in all three types of bones cells in mice and observed severe deficiencies in numerous parameters of normal bone phenotypes. STAT3 seems to play a principal role in the signaling that takes place upon mechanical loading of bone tissues and calling cells into action where they are needed. Furthermore, STAT3 has been found to be up-regulated in the early-response gene cluster following mechanical loading. Our current approach to studying STAT3’s effects on bone includes both in vivo and in vitro comparisons of WT and KO STAT3 models. The conditional knock-out of STAT3 in 8-week old mice revealed significant phenotypic variations as compared to the WT controls, while no significant differences were observed in cKO newborn pups. We also looked at immortalized WT and STAT3 KO cell lines. The STAT3 KO cells had diminished proliferation rates and decreased differentiation capabilities. Furthermore, STAT3 KO cells showed significantly reduced mRNA levels of both Wnt3a and Wnt5a when exposed to fluid shear stress. By employing available ChIP-seq data, we were able to elucidate the genome-wide binding patterns of STAT3. From the peak distribution, we can begin to uncover novel downstream effectors of STAT3 signaling that are responsible for the observed phenotypes in our conditional knockout mouse model. A preliminary look at the ChIP-seq data reveals Wnt and Nrf2 signaling to be under the putative control of STAT3. In our further research, we endeavor to experimentally confirm the ChIP-seq data for STAT3 with RNA-seq experiments in the hopes of finding potential therapeutic targets for bone pathologies.