Made available in DSpace on 2017-07-21T19:59:57Z (GMT). No. of bitstreams: 1 Nayara Carvalho Leite.pdf: 2701096 bytes, checksum: 1dca8ecc3a7945789cec9125b5a0c33b (MD5) Previous issue date: 2013-02-18 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A obesidade é um fator de risco para o desenvolvimento do diabetes tipo 2. O exercício físico reduz o tecido adiposo, modula a secreção e ação da insulina preservando a homeostase glicêmica. A administração de glutamato monossódico (MSG) induz lesões hipotalâmicas que levam a obesidade. O controle da secreção de insulina depende da formação do ATP nas células β pancreáticas, processo acoplado a rotas metabólicas glicolíticas e mitocondriais. O presente estudo investigou o efeito do exercício físico no metabolismo da glicose em ilhotas pancreáticas de ratos MSG-obesos. A obesidade foi induzida pela administração de MSG (4g/Kg). Controles (CON) receberam salina. Aos 21 dias os animais foram separados em 4 grupos CON-SED (sedentários); CON-EXE (exercitados); MSG-SED e MSG-EXE. O exercício consistiu em natação (3x/semana/30min). As ilhotas foram isoladas e incubadas com glicose (16,7 mM) na presença ou ausência dos seguintes bloqueadores do metabolismo da glicose: 1-Ácido Iodoacético (IAA, 1mM), bloqueia a glicólise; 2– Ácido alfa-ciano-4-hidroxicinâmico (-CHC, 1mM), evita o metabolismo do piruvato; 3– Fluoroacetato de Sódio (SF,2 mM) inibe o ciclo do ácido tricarboxílico (AT); 4– Rotenona (ROT, 1M) e 5- Antimicina (ANT, 50nM), inibidores respectivamente dos complexos mitocondriais I e III. A expressão proteica do transportador GLUT2 foi avaliada em ilhotas pancreáticas. Os dados foram avaliados por análises de variância (ANOVA) ou Teste t de Student (p<0,05). Ratos MSG-SED desenvolveram obesidade, resistência à insulina e hipersecreção de insulina em relação aos ratos CON-SED. A natação evitou a hiperinsulinemia, corrigiu a resistência à insulina e atenuou o excesso de tecido adiposo em ratos MSG. Ilhotas pancreáticas de ratos MSG-SED apresentam hipertrofia, aumentada expressão do GLUT2, secretando cerca de 25% mais insulina em relação a ilhotas de ratos CON-SED. Em ambos os grupos a natação reduziu em média 25% a secreção de insulina estimulada por glicose. A natação atenuou a hipertrofia dos adipócitos, das ilhotas pancreáticas, bem como corrigiu a expressão do GLUT2 em ratos MSG-obesos. O efeito do bloqueador glicolítico foi mais acentuado em ilhotas de ratos MSG-SED, indicando maior ativação desta via. O bloqueio do complexo I mitocondrial teve efeito similar entre os grupos. Todavia a inibição do complexo III foi menos acentuada em ilhotas de ratos MSG-SED. Os bloqueadores do ciclo AT e transporte do piruvato não inibiram o controle secretor de insulina em ilhotas de todos os grupos; porém o α-CHC exerceu efeito paradoxal em ilhotas de ratos MSG-SED. Alterações da glicólise; do ciclo do AT e do complexo I mitocondrial parecem não estar envolvidas na menor resposta a glicose encontrada em ilhotas de ratos exercitados. Porém, aumento da participação do complexo III mitocondrial foi observado em ilhotas de ambos os grupos exercitados. O tratamento neonatal com MSG induz obesidade, dislipidemia e resistência à insulina, eventos atenuados e/ou revertidos pela natação. A hipersecreção de insulina é corrigida pelo exercício sem alterar a via glicolítica e/ou do ciclo do AT. Todavia, a natação parece elevar a participação do complexo III mitocondrial.

O diabetes mellitus tipo 1 é uma doença autoimune causada pela destruição das células β produtoras de insulina, presentes nas ilhotas pancreáticas, por células autorreativas do sistema imune. A opção de tratamento mais utilizada são injeções diárias de insulina exógena, o que configura um tratamento não curativo. Para alcançar a independência de insulina, alternativas como o transplante de ilhotas vêm sendo estudadas. Entretanto, a disponibilidade de pâncreas de doadores cadavéricos para o isolamento destas ilhotas é pequena e os métodos de isolamento, pouco eficazes, sendo necessários de 2 a 4 doadores para atingir o número adequado de ilhotas. Além disso, o transplante apresenta problemas relacionados à enxertia, devidos principalmente à baixa vascularização, o que leva à morte de células β nos primeiros dias pós-transplante. Desta forma, estudos explorando alternativas que aumentem a sobrevivência e a funcionalidade dos transplantes e diminuam o número de ilhotas exigido por receptor fazem-se muito necessários. As células-tronco mesenquimais apresentam propriedades interessantes para aplicação em terapia celular. Entre elas, destaca-se o efeito parácrino, que exerce diversas funções benéficas, como o aumento da vascularização, nos locais onde estas células estão presentes. Sendo assim, este trabalho explorou o co-transplante de ilhotas pancreáticas com células-tronco mesenquimais derivadas de tecido adiposo, para o tratamento do diabetes mellitus em modelo murino. Os resultados mostraram que a presença destas células no grupo que recebeu o co-transplante não aumentou a taxa de cura, em relação ao grupo que recebeu somente ilhotas. No entanto, o fenômeno de reversão do diabetes foi antecipado no grupo co-transplantado, o que sugere um possível efeito angiogênico das células-tronco adiposo-derivadas presentes neste grupo. Desta forma, conclui-se que estas células podem exercer atividades benéficas, quando co-transplantadas com ilhotas pancreáticas, para o tratamento do diabetes. / Type 1 diabetes mellitus is an autoimmune disease caused by destruction of insulin-producing β cells, present in pancreatic islets, by auto-reactive cells of the immune system. The most widely used treatment option are daily injections of insulin, which configures a non-curative treatment. To achieve insulin independence, alternatives such as islet transplantation have been studied. However, the availability of pancreas from cadaveric donors for the isolation of these islets is poor and the methods for isolation, ineffective, requiring 2 to 4 donors to achieve the appropriate number of islets. In addition, transplantation presents problems related to engraftment, mainly due to poor vascularization, which leads to β cell death in the first days after transplantation. Thus, studies exploring alternatives that increase the survival and function of transplants and reduce the number of islets required by the recipient are very necessary. Mesenchymal stem cells have interesting properties for application in cell therapy. Among them is the paracrine effect, which has several beneficial functions, such as promoting vascularization in the tissues where these cells are present. Thus, the present study explored the co-transplantation of pancreatic islets with mesenchymal stem cells derived from adipose tissue for the treatment of diabetes mellitus in mice. The results showed that the presence of these cells in the group that received co-transplantation did not increase the cure rate, compared to the group that received islets alone. However, the phenomenon of diabetes reversion was anticipated in co-transplanted animals, which suggests a possible angiogenic effect of adipose-derived stem cells present in this group. Thus, we conclude that these cells may exert beneficial functions when co-transplanted with pancreatic islets for the treatment of diabetes.

Notre travail de thèse avait pour objet l’étude du potentiel anti-diabétique des urolithines A, B, C et D, métabolites de polyphénols formés par le microbiote colique à partir des tanins de l’acide ellagique (présents notamment dans la grenade et les noix).La première partie, bibliographique, constitue un rappel :• de la régulation de l’équilibre glycémique et le rôle de la sécrétion d’insuline dans cette régulation ; •de l ‘épidémiologie et la physiopathologie du diabète type 2 (DT2) ; •des polyphénols et leurs métabolites, ainsi que de leurs effets antidiabétiques potentiels.La seconde partie décrit les effets des urolithines sur différents modèles expérimentaux : •Sur un modèle de cellules β insulino-sécrétrices (lignée INS-1), les urolithines induisent une amplification concentration-dépendante de la sécrétion d’insuline induite par le glucose, mais également par d’autres sécrétagogues comme un analogue du GLP-1 ou une sulfonylurée (médicaments utilisés dans le diabète). Les urolithines préviennent également l’altération sécrétoire induite par un stress oxydant. •L’effet insulino-sécrétoire des urolithines a été confirmé sur îlots de Langerhans isolés. •L’urolithine C étant apparu comme le composé le plus prometteur, nous avons poursuivi la caractérisation de son activité sur un modèle ex vivo mimant la situation physiologique, le pancréas isolé perfusé. Alors que l’effet sécrétoire de l’urolithine C n’apparaît pas en présence de 5mM de glucose, l’urolithine C (20µM) a stimulé la sécrétion d’insuline dans des conditions de stimulation modérée de la sécrétion d’insuline par le glucose (8.3mM). Cet effet est strictement dépendant du glucose, la sécrétion d’insuline retournant immédiatement à son niveau basal lors du passage de 8,3 à 5mM de glucose en présence d’urolithine C. •Des études de pharmacocinétique ont permis de mettre au point une méthodologie de dosage plasmatique de l’urolithine C dans le plasma de rat par chromatographie liquide / ionisation electrospray /spectrographie de masse en tandem. Cette méthodologie a été appliquée à une première étude pharmacocinétique chez le rat après injection de 10mg/kg d’urolithine C par voie intra-péritonéale. Cette étude montre notamment que le profil pharmacocinétique suit un modèle à 3 compartiments et suggère un stockage tissulaire du composé.D’autres résultats (confidentiels) ne peuvent être évoqués dans ce résumé mais confirment l’intérêt potentiel de l’urolithine C dans le traitement du diabète de type 2 en tant que médicament insulinotrope dépendant du glucose. / The objective of our thesis was to study the anti-diabetic potential of metabolites of ellagic acid tanins, present notably in pomegranate and nuts, that are formed by the colon microbiote. The metabolites are urolithins A, B, C and D.The first part of thesis is bibliographic and reviews: •The control of glycemic plasma levels, and in particular the role of insulin secretion in this process; • The pathophysiology of Type 2 Diabetes (T2D); •The various polyphenols and their metabolites, along with their potential anti-diabetic activity.The second part describes the effects of urolithins on various experimental models: •On a model of insulin secreting beta cells (the INS-1cell line), urolithins concentration-dependently amplified insulin secretion induced by glucose, but also by insulinotropic drugs used in the treatment of T2D such as a GLP-1 analogue or a sulfonylurea. In addition, urolithins were able to induce insulin secretion on cells rendered unresponsive to glucose by oxidative stress. • The insulinotropic effect of urolithins was also confirmed on isolated rat islets of Langerhans. •As urolithin C appeared to be the most promising antidiabetic compound, we further characterized its activity on an ex vivo model mimicking the physiological situation, the isolated infused pancreas. While urolithin C (20µM) had no effect in the presence of 5 mM glucose concentration, it amplified the stimulation of insulin secretion in the presence of 8.3mM glucose. The effect of urolithin C was also strictly glucose-dependent, as insulin secretion immediately returned to basal level when glucose concentration was switched from 8.3 to 5mM glucose in the presence of urolithin C. •We also conducted studies aiming at designing a validated methodology for rat plasma urolithin C determination using a liquid chromatography-electrospray ionization-tandem mass spectrometry method. The applicability of this assay was demonstrated in a preclinical pharmacokinetic study carried out in rats receiving intraperitoneal administration of urolithin C (10mg/kg). We found that the urolithin C followed a three-compartment model, suggesting a long-term tissue storage of urolithin C.Some other (confidential) results, not described in this abstract, confirmed urolithin C as a potential glucose-dependent insulinotropic treatment for type 2 diabetes.

Another growth factor candidate is a recently recognized bioactive peptide, islet-neogenesis associated protein (INGAP). A master pancreatic transcription factor, pancreatic duodenal homeobox-1 (Pdx-1), was overexpressed in PSCs by the adenovirus-mediated transfer method in the present study. With the infection of adenovirus expressing Pdx-1, several beta-cell developmental genes, including Isl-1, Beta2, Nkx2.2, Nkx6.1 and the endogenous Pdx-1 were found to be upregulated temporally in our PSCs-derived ICCs. Meanwhile, previous study has shown that Pdx-1/INGAP-positive cells represent a new stem cell subpopulation during early stage of pancreatic development. We thus explore whether any functional integration of Pdx-1 and INGAP in the growth and functional maturation of PSCs. In order to achieve this proposition, the effects of over-expressing PSCs with the Pdx-1 adenovirus in conjunction with the treatment of INGAP were then investigated. Interestingly, differentiation of the PSC-derived ICCs was not further enhanced by the synergistic treatment of Pdx-1 and INGAP when compared to those ICCs infected with adenovirus expressing Pdx-1 alone, as revealed by the endogenous Pdx-1 and insulin gene expression and their C-peptide content. These data might provide some clues to the intricate interaction between Pdx-1 and INGAP in regulating the ICC and/or the pancreatic endocrine differentiation. (Abstract shortened by UMI.) / Due to the scarcity of fetal pancreas for generating functional insulin-secreting cell clusters for sufficient islet transplantation, we targeted for searching pancreatic stem/progenitor cells. Putative PSCs can be aggregated and differentiated into islet-like cell clusters (ICCs) when exposed to serum-free medium containing various conventional growth factors, including HGF, GLP-1, betacellulin and nicotinamide. / Fetal pancreatic tissue consisting of immature progenitor cells serves as a potential source of stem cells as they possess a higher replicative capacity and longevity than their adult counterparts. / Two novel candidates and a key pancreatic transcription factor on the PSC/ICC proliferation and differentiation were investigated in the present study. One of them is a ubiquitously expressed multi-PDZ-domain protein, PDZ-domain-containing 2 (PDZD2), which was previously found to express in the mouse beta cells and exhibit mitogenic effects in beta cell line. Results showed that PDZD2 was detected in high levels in both human fetal pancreas and in PSCs. Results indicate the potential involvement of PDZD2 in regulating PSCs proliferation and differentiation and pancreatic development. / Suen Po Man, Ada. / "July 2007." / Adviser: P.S. Leung. / Source: Dissertation Abstracts International, Volume: 69-01, Section: B, page: 0051. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 194-214). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Malignancy within the pancreatic head can arise from pancreatic duct, distal bile duct, ampulla or duodenum. Since September 2000, surgery for all pancreatic head malignancy (PHM) has been centralised into regional pancreatic centres where assessment of preoperative imaging and subsequent surgery is undertaken. As part of this guidance, surgery must be performed within 62-days of referral. This project will assess four aspects of the pre-operative pathway in PHM: 1) Potential variation in outcome of patients referred from different sites within a Cancer Network 2) Potential variation in outcome associated with different intervals to surgery within the 62 day guideline 3) The ability of interpretation of heterogeneous pre-operative CT scans from different hospitals to determine the resectability of PHM 4) The ability of CT scan to distinguish the different tumour types of PHM Images of a consecutive series of patients were re-reported and compared with final pathology reports. Good agreement was noted in determining the tumour origin of PHM (observed agreement = 0.758, Kappa= 0.6 (0.51-0.68)). In the assessment surgical outcomes, geographical isolation from the regional centre was not associated with delay to surgery. Variation in outcome between referral centres was however noted but this was not associated with travel distance. Although little association was noted between delay to surgery and outcome overall, a paradoxical improvement in survival was noted however for the small group of patients with ampullary tumours who waited longer than the median interval to surgery.

Le rôle des acides biliaires a évolué ces dernières années passant de simples molécules solubilisatrices des lipides à des composés à activité métabolique. En plus de leur fonction dans l’absorption des lipides post-repas, ils ont été montrés comme stimulant de nombreuses voies de signalisation modulant l’expression de gènes clefs du métabolisme et de nombreux mécanismes physiologiques via l’activation de récepteurs spécifiques tels que les récepteurs « Farnesoid X receptor » (FXR) et le récepteur membranaire couplé à une protéine G, TGR5. La protéine TGR5 codée par le gène GPBAR1, aussi connue sous le nom de « G-protein-membrane-type receptor for bile acids » (M-BAR) est le premier récepteur couplé à une protéine G spécifique aux acides biliaires ayant été mis en évidence. Cette protéine est exprimée dans de nombreux tissus clefs du métabolisme énergétique tels que les cellules L intestinales, le tissu adipeux, les reins, le muscle squelettique et le pancréas. En réponse à la fixation des acides biliaires au récepteur TGR5, celui-ci va être internalisé et sa sous-unité G&#945;S va être libérée. Ce mécanisme va ensuite activer l’adénylate cyclase et augmenter la production d’AMPc à l’origine de l’activation des voies de signalisations liées à la protéine kinase A (PKA). Une fois activée, la PKA va induire la phosphorylation des protéines « cAMP-response element-binding » (CREB) et permettre la modulation de l’expression de gènes cibles.Ces dernières années de nombreux travaux ont eu pour but d’étudier le rôle du récepteur TGR5 dans le métabolisme. Chez la souris, l’activation du récepteur TGR5 stimule la dépense énergétique dans le tissu adipeux brun et dans le muscle squelettique et prévient le développement de l’obésité et de l’insulino-résistance induites par un régime riche en graisses. Le récepteur TGR5 est également impliqué au niveau des cellules L intestinales sécrétrices du GLP-1. Il y joue un rôle essentiel dans l’homéostasie glucidique via la régulation de l’activité pancréatique, des sécrétions de l’insuline et du glucagon, de l’inhibition de la vidange gastrique ou encore de la modulation des messages de satiété via des voies neuroendocrines. TGR5 présente également des fonctions immunologiques avec une expression connue dans les cellules de l’immunité telles que les monocytes, les macrophages alvéolaires ou encore les cellules de Kupffer. TGR5 a également été mis en évidence comme régulateur des mécanismes d’inflammations via les macrophages avec une diminution de l’expression des cytokines pro-inflammatoires. A l’opposé, l’activation de TGR5 serait impliquée dans de nombreux processus pathologiques tels que, le développement de carcinomes gastro-intestinaux, les pancréatites, la lithiase biliaire, suggérant un rôle potentiel du récepteur TGR5 dans la régulation de voies de signalisation responsables de la prolifération et de la mort cellulaire [...] / Bile acids (BAs) have evolved over the years from being considered as simple lipid solubilizers to metabolically active molecules. In addition to their function in dietary lipid absorption, they have also been shown to activate farnesoid X receptor (FXR) and TGR5 receptors to initiate signaling pathways and regulate metabolic gene transcription. TGR5 (encoded by the GPBAR1 gene), also known as G-protein-membrane-type receptor for bile acids (M-BAR) or G-protein-coupled bile acid receptor 1 (GPBAR1), was the first identified G-protein coupled receptor specific for bile acids. In normal individuals, the highest level of GPBAR1 mRNA expression was reported in the gallbladder, placenta and spleen, followed by moderate expression in other tissues including lungs, liver, stomach, small intestine and adipose tissue, with a relatively low level of expression in kidney, skeletal muscles and pancreas. In response to binding of BAs to the ligand-binding pocket of the TGR5 protein, the TGR5 receptor is internalized and the G&#945;S subunit is released. This mechanism leads to activation of adenylate cyclase and an increase in cAMP production resulting in induction of the protein kinase A (PKA) pathway. Subsequently, PKA phosphorylates the cAMP-response element-binding protein (CREB) and enhances the transcription of its target genes in response to extracellular signals.To date, extensive work has been done to investigate the role of TGR5 in metabolism. In rodents, BA-activated TGR5 receptor stimulates energy expenditure in brown adipose tissue and skeletal muscle and prevents obesity and insulin resistance induced by a high fat diet. TGR5 is also implicated in intestinal L-cells secreted GLP-1, which plays an essential role in glucose homeostasis through the stimulation of glucose-dependent-insulin-secretion and inhibition of glucagon secretion, inhibition of gastric emptying and increasing satiety through neuroendocrine pathways. In terms of the immunological function of TGR5, it is now known that TGR5 is expressed in several immune cells such as monocytes, alveolar macrophages and Kupffer cells. The beneficial effects of TGR5 on macrophage-driven inflammation include reduced proinflammatory cytokine expression, thus protecting against atherosclerosis and liver steatosis. On the contrary, TGR5 activation has also been implicated in itch and analgesia, gastrointestinal-tract cell carcinogenesis, pancreatitis, and cholelithiasis, suggesting a potential role for TGR5 as a regulator of signal transduction pathways responsible for cell proliferation and apoptosis. BAs may also influence islet function via both direct and indirect mechanisms as recent studies have shown that Farnesoid X receptor (FXR) is expressed by pancreatic beta cells, and regulates insulin signaling in cultured cell lines. Kumar et al., [14] also reported that the TGR5 agonists INT-777 + oleanolic acid (OA) stimulated glucose-mediated insulin release via TGR5 activation, also in cultured cells. Still, little is known about the regulation of TGR5 expression or its involvement in pancreatic hormone secretion in response to physiological or pathological conditions such as T2D, as these studies have been performed mainly in cultured cell lines. In these contexts, the biological function of TGR5 remains enigmatic. The aim of the present study was first to establish the specific expression of TGR5 in human pancreatic islet cell subtypes. Then, a cross-sectional cohort of human islets isolated from individuals with various degrees of insulin resistance was exploited to determine if TGR5 expression and function was modified in T2D. Finally to determine if targeting TGR5 is clinically relevant, human islets were treated in-vitro with a specific agonist of TGR5 or with siRNA directed against TGR5 and hormone secretion assessed to establish whether TGR5 activation or inhibition modulate pancreatic hormone secretion.

Introdução: Os benefícios do emprego do selante de fibrina no reforço de anastomoses pancreatico-jejunais, após ressecção duodenopancreática, visando a redução da incidência de fístula pancreática pós operatória (FPPO), ainda são questionáveis. Objetivo: Avaliar a influência do emprego do selante de fibrina na anastomose pancreatico-jejunal, após duodenopancreatectomia, na incidência de fístula, bem como suas consequências clínicas e os custos hospitalares. Metodologia: Estudo retrospectivo de 62 pacientes consecutivos submetidos a duodenopancreatectomia, divididos em dois grupos: 31 pacientes utilizando o selante de fibrina (GCS) e 31 pacientes sem o emprego de selante (GSS). As variáveis estudadas foram agrupadas em epidemiológicas, clínicas, laboratoriais, com destaque para a incidência de fístula pancreática, classificada segundo a definição do International Study Group on Pancreatic Fistula, suas complicações pós operatórias catalogadas segundo a classificação de Clavien e suas repercussões na assistência e nos seus custos avaliados pelo método de absorção com rateio simples de todas as despesas, exceto a despesa com medicamentos, tratada de forma separada. Resultados: Os grupos foram homogêneos para os parâmetros epidemiológicos, clínicos, e laboratoriais e não foram registradas diferenças significativas na comparação da evolução pós operatória e dos indicadores assistenciais hospitalares. Por outro lado, os custos hospitalares foram mais elevados no GCS, em relação ao GSS (p<0,0001). Conclusão: O emprego do selante de fibrina, no reforço da anastomose pancreatico-jejunal, em pacientes submetidos a duodenopancreatectomias, nas condições estudadas, não melhorou os resultados clínicos e assistenciais e ainda aumentou os custos hospitalares. / Introduction: The benefits of fibrin sealant employment in strengthening pancreatico-jejunal anastomosis after duodenopancreatic resection, reducing the incidence of pancreatic fistula postoperative (PFPO) are still questionable. Objective: To evaluate the influence of the use of fibrin sealant in pancreatico-jejunal anastomosis after pancreaticoduodenectomy in the incidence of fistula and its clinical consequences and hospital costs. Methodology: A retrospective study of 62 consecutive patients who underwent pancreaticoduodenectomy, divided into two groups: 31 patients using fibrin sealant (GCS) and 31 patients without the sealant employment (GSS). The variables were grouped into epidemiological, clinical, laboratory, especially the incidence of pancreatic fistula classified as defined by the International Study Group on Pancreatic Fistula, their postoperative complications cataloged according to Clavien rating and its repercussions on care and its costs assessed by the absorption method with simple apportionment of all expenses except the expenditure on medicines, treated separately. Results: The groups were homogeneous for clinical, epidemiological and laboratory parameters and no significant differences were recorded in the comparison given postoperative progress and hospital assistance indicators. Moreover, hospital costs were higher in GCS, with respect to GSS (p <0.0001). Conclusion: The use of fibrin sealant in pancreatojejunal anastomosis after pancreaticoduodenectomy, in the studied conditions, did not improve the results of care and also increased hospital costs

Ma, Man Ting. / "July 2010." / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 186-207). / Abstracts in English and Chinese. / Abstract --- p.I / List of Publications --- p.VI / Acknowledgements --- p.VIII / Table of Contents --- p.X / List of Figures --- p.XV / List of Tables --- p.XVIII / List of Abbreviations --- p.XIX / Chapter CHAPTER1 --- INTRODUCTION / Chapter 1.1 --- The Pancreas --- p.2 / Chapter 1.1.1 --- Structure of pancreas --- p.2 / Chapter 1.1.2 --- Structure and function of exocrine pancreas --- p.6 / Chapter 1.1.3 --- Structure and function of endocrine pancreas --- p.9 / Chapter 1.1.3.1 --- Pancreatic islet and islet cells --- p.9 / Chapter 1.1.3.2 --- Glucose-stimulated insulin secretion from islets --- p.12 / Chapter 1.2 --- Type 1 Diabetes Mellitus (T1DM) --- p.14 / Chapter 1.2.1 --- Pathophysiology of Diabetes Mellitus --- p.14 / Chapter 1.2.2 --- Autoimmunity in T1DM --- p.17 / Chapter 1.2.3 --- Management ofTlDM --- p.20 / Chapter 1.2.3.1 --- Insulin replacement --- p.20 / Chapter 1.2.3.2 --- Pancreas and islet transplantation --- p.21 / Chapter 1.2.3.3 --- Stem-cell-based transplantation --- p.22 / Chapter 1.3 --- The Adaptive Immune System --- p.26 / Chapter 1.3.1 --- T-lymphocytes --- p.26 / Chapter 1.3.2 --- B-lymphocytes --- p.29 / Chapter 1.3.3 --- Major histocompatibility complex (MHC) --- p.30 / Chapter 1.3.3.1 --- Classification of MHC molecules --- p.30 / Chapter 1.3.3.2 --- Structure of MHC class I and II molecules --- p.32 / Chapter 1.3.3.3 --- Function and regulation of MHC molecules --- p.34 / Chapter 1.3.4 --- HLA-G and its immuno-modulatory properties --- p.36 / Chapter 1.4 --- Transplantation Rejection --- p.40 / Chapter 1.4.1 --- Mechanisms involved in transplantation rejection --- p.40 / Chapter 1.4.2 --- Immunobiology of rejection --- p.41 / Chapter 1.4.2.1 --- Direct allorecognition pathway --- p.42 / Chapter 1.4.2.2 --- Indirect allorecognition pathway --- p.43 / Chapter 1.4.2.3 --- Semi-direct allorecognition pathway --- p.43 / Chapter 1.4.3 --- Xenotransplantation --- p.46 / Chapter 1.5 --- Cytokines and Immunity --- p.48 / Chapter 1.5.1 --- Interferons --- p.48 / Chapter 1.5.1.1 --- Interferon-γ and its immune regulation --- p.49 / Chapter 1.5.1.2 --- Effect and kinetics of interferon-γ on MHC molecules expression --- p.53 / Chapter 1.5.1.3 --- Regulation of interferon-γ production --- p.56 / Chapter 1.5.2 --- Interlukins --- p.58 / Chapter 1.5.2.1 --- IL-10 and its immune regulation --- p.58 / Chapter 1.5.2.2 --- IL-10 and HLA-G --- p.59 / Chapter 1.6 --- Stem Cells and their Immunogenicity --- p.62 / Chapter 1.6.1 --- Embroynic stem cells --- p.62 / Chapter 1.6.2 --- Mesenchymal stem cells --- p.64 / Chapter 1.6.3 --- Neural stem cells --- p.68 / Chapter 1.6.4 --- Fetal stem cells --- p.69 / Chapter 1.6.5 --- Potential immuno-study in human fetal pancreatic stem cells --- p.70 / Chapter 1.7 --- Aims and Objectives of study --- p.72 / Chapter CHAPTER2 --- MATERIALS AND METHODS / Chapter 2.1 --- Isolation of Pancreatic Progenitors (PPCs) from Human Fetal Pancreas and Induction of Islet-like Cell Cluster (ICCs) Differentiation --- p.75 / Chapter 2.1.1 --- Tissue procurement --- p.75 / Chapter 2.1.2 --- Tissue processing and PPCs culture --- p.75 / Chapter 2.1.3 --- In vitro differentiation of PPCs into ICCs --- p.78 / Chapter 2.1.4 --- Interferon-γ and IL-10 treatment --- p.80 / Chapter 2.2 --- Cell culture of human placental Choriocarcinoma JEG-3 Cell Line --- p.81 / Chapter 2.3 --- RNA Expression Detection --- p.82 / Chapter 2.3.1 --- RNA isolation --- p.82 / Chapter 2.3.2 --- Reverse transcriptase (RT) --- p.83 / Chapter 2.3.3 --- Design of primers for Polymerase Chain Reaction (PCR) and Real-time PCR --- p.84 / Chapter 2.3.4 --- PCR --- p.86 / Chapter 2.3.5 --- Real-time PCR analysis --- p.88 / Chapter 2.3.6 --- Calculation using the comparative CT method --- p.90 / Chapter 2.4 --- Flow Cytometry --- p.91 / Chapter 2.5 --- Western Blotting Analysis --- p.93 / Chapter 2.5.1 --- Protein extraction and quantification --- p.93 / Chapter 2.5.2 --- Western blotting --- p.93 / Chapter 2.6 --- Mixed Lymphocyte Reaction (MLR) --- p.95 / Chapter 2.6.1 --- Isolation of peripheral blood mononuclear cells (PBMCs) --- p.95 / Chapter 2.6.2 --- PPC-PBMCs MLR --- p.98 / Chapter 2.6.3 --- ICC-PBMCs MLR --- p.98 / Chapter 2.6.4 --- Proliferation assay --- p.99 / Chapter 2.7 --- ICC Transplantation --- p.101 / Chapter 2.7.1 --- Streptozotocin-induced diabetic animals for transplantation --- p.101 / Chapter 2.7.2 --- Procedures of ICCs transplantation --- p.102 / Chapter 2.8 --- Histological Analysis of ICC Graft --- p.105 / Chapter 2.8.1 --- H&E staining --- p.105 / Chapter 2.8.2 --- DAB staining --- p.106 / Chapter 2.8.3 --- Immunofluorescence staining --- p.107 / Chapter 2.9 --- Enzyme-linked Immunosorbent Assay (ELISA) --- p.109 / Chapter 2.10 --- Statistical Data Analysis --- p.110 / Chapter CHAPTER3 --- RESULTS / Chapter 3.1 --- Immuno-characterization of PPCs and ICCs --- p.112 / Chapter 3.2 --- Effect of cytokines on immune-properties of PPCs and ICCs --- p.115 / Chapter 3.2.1 --- Effect of lFN-γ on MHC-I expression in PPCs --- p.115 / Chapter 3.2.2 --- Effect of lFN-γ and IL-10 on HLA-G expression in PPCs and ICCs --- p.119 / Chapter 3.2.3 --- Effect of IFN-γ on B7H4 expression in PPCs --- p.123 / Chapter 3.3 --- Comparison of immune-properties of PPCs and ICCs from 1st and 2nd trimester --- p.125 / Chapter 3.3.1 --- Differential expression of MHC molecules in PPCs --- p.125 / Chapter 3.3.2 --- Different immune-related gene expression in PPCs and ICCs --- p.128 / Chapter 3.3.3 --- Comparison of IFN-γ activated MHC molecules expression in PPCs/ICCs --- p.134 / Chapter 3.3.4 --- Comparison of other IFN-γ activated genes expression in PPCs --- p.139 / Chapter 3.4 --- Mixed lymphocyte reaction of PPCs from 1st and 2nd trimester --- p.143 / Chapter 3.4.1 --- Effect of PPCs on proliferation of PBMC --- p.143 / Chapter 3.4.2 --- Effect of ICCs on proliferation of PBMC --- p.145 / Chapter 3.4.3 --- Effect of PPCs on cytokine production in PBMC --- p.149 / Chapter 3.5 --- Xenotransplantation of ICCs into diabetic mouse model --- p.152 / Chapter 3.5.1 --- Blood glucose level of diabetic mice after transplantation --- p.152 / Chapter 3.5.2 --- Histological evaluation of transplanted ICCs grafts --- p.154 / Chapter 3.5.3 --- Infiltration of CD45 into transplanted grafts of 1st and 2nd trimester --- p.158 / Chapter CHAPTER4 --- DISCUSSION / Chapter 4.1 --- Expression of selected immuno-regulated genes in PPCs and ICCs --- p.163 / Chapter 4.2 --- Effect of IFN-g and IL-10 on expression of immuno-regulated genes in PPCs and ICCs --- p.166 / Chapter 4.3 --- In vitro studies on immunogenicity of PPCs and ICCs from first and second trimester --- p.171 / Chapter 4.3.1 --- Immune-related genes expression --- p.171 / Chapter 4.3.2 --- IFN-γ activated gene expression --- p.173 / Chapter 4.3.3 --- Mixed lymphocyte reaction --- p.175 / Chapter 4.3.4 --- Cytokine production of PBMC in MLR --- p.179 / Chapter 4.4 --- In vivo Xenotransplantation of ICCs into diabetic mouse model --- p.181 / Chapter 4.5 --- Conclusion --- p.187 / Chapter 4.6 --- Further studies --- p.188 / Chapter CHAPTER5 --- BIBLIOGRAPHY / Bibliography by Alphabetical Order --- p.189

Ng, Ka Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 179-204). / Abstracts in English and Chinese. / Abstract --- p.I / 摘要 --- p.IV / Publications --- p.VII / Acknowledgements --- p.VIII / Table of contents --- p.IX / List of figures --- p.XV / List of tables --- p.XVII / List of abbreviations --- p.XVIII / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- The Pancreas --- p.2 / Chapter 1.1.1 --- Anatomy of Pancreas --- p.2 / Chapter 1.1.2 --- The Exocrine Pancreas --- p.4 / Chapter 1.1.3 --- The Endocrine Pancreas --- p.5 / Chapter 1.1.3.1 --- Structure of Islets --- p.5 / Chapter 1.1.3.2 --- "Functions of α-, β-, y-, ð-, Σ-and PP-cells in Islets" --- p.7 / Chapter 1.1.4 --- Overview of Pancreas Development --- p.9 / Chapter 1.1.4.1 --- Organ Morphology --- p.10 / Chapter 1.1.4.2 --- Cyto-differentiation --- p.12 / Chapter 1.1.4.3 --- Control by Transcriptional Factors --- p.14 / Chapter 1.1.5 --- Postnatal Pancreas Development and Regeneration --- p.18 / Chapter 1.1.5.1 --- Proliferation of Pre-existing β-cells --- p.19 / Chapter 1.1.5.2 --- Neogenesis from Precursor Cells --- p.20 / Chapter 1.1.5.3 --- Transdifferentiation of other Cells --- p.20 / Chapter 1.2 --- Diabetes Mellitus --- p.22 / Chapter 1.2.1 --- Pathophysiology of Diabetes Mellitus and Current Treatments --- p.24 / Chapter 1.2.1.1 --- Type I Diabetes Mellitus --- p.24 / Chapter 1.2.1.2 --- Type II Diabetes Mellitus --- p.25 / Chapter 1.2.1.3 --- Gestational Diabetes --- p.27 / Chapter 1.2.1.4 --- Secondary Diabetes --- p.28 / Chapter 1.3 --- Stem Cell therapy --- p.29 / Chapter 1.3.1 --- Stem Cell --- p.29 / Chapter 1.3.1.1 --- Mesenchymal Stem Sell --- p.31 / Chapter 1.3.1.2 --- Embryonic Stem Cell --- p.35 / Chapter 1.3.1.3 --- Induced Pluripotent Stem Cell --- p.36 / Chapter 1.3.2 --- Islets Engineering --- p.37 / Chapter 1.3.2.1 --- Genetic Modification --- p.37 / Chapter 1.3.2.2 --- Directed Differentiation --- p.38 / Chapter 1.3.2.3 --- Microenvironment --- p.38 / Chapter 1.3.2.4 --- In vivo Regeneration --- p.39 / Chapter 1.3.2.5 --- Cell Fusions --- p.40 / Chapter 1.3.2.6 --- Combinatory Treatments --- p.40 / Chapter 1.4 --- The Vitamin A & Vitamin D System --- p.42 / Chapter 1.4.1 --- The Vitamin A --- p.42 / Chapter 1.4.2 --- Vitamin A Metabolism --- p.44 / Chapter 1.4.3 --- Roles of vitamin A in Pancreatic Development --- p.46 / Chapter 1.4.4 --- The Vitamin D --- p.48 / Chapter 1.4.5 --- Vitamin D Metabolism --- p.49 / Chapter 1.4.6 --- Metabolic Functions of Vitamin D in Islets --- p.51 / Chapter 1.4.7 --- Cod Liver Oil --- p.53 / Chapter 1.4.8 --- Interactions between Vitamin A and Vitamin D --- p.53 / Chapter 1.5 --- The Relations of Liver and Pancreas Development --- p.55 / Chapter 1.5.1 --- Endoderm Induction for Hepatic and Pancreatic Differentiation of ESCs --- p.55 / Chapter 1.5.2 --- Bipotential Precursor Population within Embryonic Endoderm --- p.56 / Chapter 1.5.3 --- Pancreatic Islets Promote Mature Liver Hepatocytes Proliferation --- p.57 / Chapter 1.5.4 --- Transdifferentiation --- p.57 / Chapter 1.5.5 --- Transplantation in Liver Niche Promotes Maturation of Insulin-Producing Cells --- p.60 / Chapter 1.5.6 --- Neuronal Relay from the Liver to Pancreatic --- p.61 / Chapter 1.5.7 --- Development of Islets in the Nile Tilapia --- p.62 / Chapter 1.6 --- The Insulin-like Growth Factor-I (IGF1) --- p.64 / Chapter 1.6.1 --- IGF1 System --- p.64 / Chapter 1.6.2 --- IGF 1 Regulation --- p.65 / Chapter 1.6.3 --- Roles of IGF 1 in Pancreatic Development and Regeneration --- p.68 / Chapter 1.7 --- Aims and Objectives of Study --- p.70 / Chapter Chapter 2 --- General Materials and Methods / Chapter 2.1 --- Pancreatic progenitor cells (PPCs) and liver stromal cells (LSCs) isolation and cell culture --- p.72 / Chapter 2.1.1 --- Tissue procurement --- p.72 / Chapter 2.1.2 --- PPC and LSC culture --- p.72 / Chapter 2.1.3 --- "Treatments of vitamin A, vitamin D and IGF 1" --- p.76 / Chapter 2.1.4 --- "Cell culture of Caco-2, HepG2 and DU-145" --- p.76 / Chapter 2.2 --- Induction of Islet-like Cell Clusters (ICCs) Differentiation --- p.77 / Chapter 2.2.1 --- In vitro Directed Differentiation --- p.77 / Chapter 2.2.2 --- In vitro LSC Microenvironment --- p.77 / Chapter 2.3 --- RNA Expression Detection --- p.79 / Chapter 2.3.1 --- RNA isolation --- p.79 / Chapter 2.3.2 --- Reverse Transcription --- p.79 / Chapter 2.3.3 --- Polymerase Chain Reaction (PCR) --- p.80 / Chapter 2.3.4 --- Realtime PCR --- p.81 / Chapter 2.4 --- Immunocytochemistry --- p.83 / Chapter 2.5 --- Western Blotting --- p.85 / Chapter 2.5.1 --- Protein extraction and quantification --- p.85 / Chapter 2.5.2 --- Western Blotting --- p.85 / Chapter 2.6 --- Enzyme-linked Immunosorbent Assay (ELISA) --- p.87 / Chapter 2.6.1 --- Detection of cell viability --- p.87 / Chapter 2.6.2 --- Detection of cell proliferation --- p.87 / Chapter 2.6.3 --- Measurement of Cell death --- p.88 / Chapter 2.6.4 --- Measurement of IGF 1 level in condition medium --- p.89 / Chapter 2.6.5 --- Measurement of glucose induced insulin secretion --- p.90 / Chapter 2.7 --- Regeneration model --- p.92 / Chapter 2.7.1 --- Regeneration model in neonatal-STZ rat --- p.92 / Chapter 2.7.2 --- Change in IGF1 expression in pancreas and liver --- p.92 / Chapter 2.8 --- Statistical Data Analysis --- p.93 / Chapter Chapter 3 --- Vitamin D and vitamin A receptor expression and the proliferative effects of ligand activation of these receptors on the development of pancreatic progenitor cells derived from human fetal pancreas. (Stem Cell Rev. 2011;7:53-63) / Chapter 3.1 --- Abstract --- p.95 / Chapter 3.2 --- Introduction --- p.97 / Chapter 3.3 --- Materials and Methods --- p.101 / Chapter 3.3.1 --- Fetal Tissue Procurement --- p.101 / Chapter 3.3.2 --- Culture of Pancreatic Progenitor Cells --- p.101 / Chapter 3.3.3 --- RNA Expression Analysis by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) --- p.102 / Chapter 3.3.4 --- Western Blot Analysis --- p.103 / Chapter 3.3.5 --- Immunocytochemstry --- p.105 / Chapter 3.3.6 --- PPC Proliferation Assays --- p.106 / Chapter 3.3.7 --- PPC Cell Death Assays --- p.107 / Chapter 3.3.8 --- Statistical Data Analysis --- p.108 / Chapter 3.4 --- Results --- p.110 / Chapter 3.4.1 --- "Expression and Localization of RAR, VDR and RXR, CYP26 and CYP24 in PPCs" --- p.110 / Chapter 3.4.2 --- Incubation of PPC with atRA Enhances PPC Viability due to Increased Proliferation and Anti-apoptosis --- p.111 / Chapter 3.4.3 --- Incubation of PPCs with Calcitriol Enhances Viability due to Increased Proliferation --- p.111 / Chapter 3.4.4 --- Both atRA and Calcitriol Induce Up-regulation of both the RAR and the VDR but not the RXR --- p.112 / Chapter 3.4.5 --- Combination Treatment with atRA and Calcitriol on Cell Viability and NGN3 Expression --- p.112 / Chapter 3.5 --- Discussion --- p.114 / Chapter Chapter 4 --- Human fetal liver stromal cell co-culture enhances the growth and differentiation of pancreatic progenitor cells into islet-like cell clusters (In submission to Gastroenterology) / Chapter 4.1 --- Abstract --- p.128 / Chapter 4.2 --- Introduction --- p.129 / Chapter 4.3 --- Materials and Methods --- p.133 / Chapter 4.3.1 --- Use of human and animal tissues --- p.133 / Chapter 4.3.2 --- "Cell preparation, characterizations and Differentiation" --- p.133 / Chapter 4.3.3 --- Examination of PPC growth and ICC differentiation and functions with LSC co-culture --- p.133 / Chapter 4.3.3 --- Identification of growth factors and investigation of their effects --- p.134 / Chapter 4.3.4 --- Statistical Analysis --- p.135 / Chapter 4.4 --- Results --- p.136 / Chapter 4.4.1 --- "Isolation, Culture and Characterizations of LSCs" --- p.136 / Chapter 4.4.2 --- Establishment of LSC co-culture system --- p.136 / Chapter 4.4.3 --- LSC co-culture enhances PPC-derived ICC differentiation --- p.137 / Chapter 4.4.4 --- Differential expression of mRNA for cytokines and growth factors between 1st and 2nd trimester LSCs --- p.138 / Chapter 4.4.5 --- Characterization of IGF 1 receptors in PPCs and the effects of exogenous IGF1 on PPC growth and ICC differentiation --- p.139 / Chapter 4.4.6 --- Neutralizing antibodies against IGF1R inhibit ICC differentiation --- p.140 / Chapter 4.5 --- Discussion --- p.142 / Chapter 4.6 --- Supplementary Materials and Methods --- p.147 / Chapter 4.6.1 --- Cell Preparation and culture --- p.147 / Chapter 4.6.2 --- In Vitro ICC differentiation --- p.148 / Chapter 4.6.3 --- RNA expression analysis --- p.149 / Chapter 4.6.4 --- Immunocytochemistry --- p.149 / Chapter 4.6.5 --- PPC viability and cell count assays --- p.150 / Chapter 4.6.6 --- IGF1 and insulin ELISA --- p.151 / Chapter 4.6.7 --- Western blotting analysis --- p.152 / Chapter 4.6.8 --- Neonatal streptozotocin regeneration model --- p.153 / Chapter Chapter 5 --- General Discussion and Future Studies / Chapter 5.1 --- General Discussion --- p.165 / Chapter 5.1.1 --- Proliferative effects and enhance expression of NGN3 by vitamin A and vitamin D on PPC --- p.166 / Chapter 5.1.2 --- Induction of PPC derived ICCs by LSCs --- p.169 / Chapter 5.1.3 --- Potential effects of liver stroma derived IGF1 on PPC derived ICCs differentiation --- p.172 / Chapter 5.1.4 --- Significance of islet engineering in the management of diabetes --- p.174 / Chapter 5.1.5 --- Conclusions --- p.176 / Chapter 5.2 --- Future Studies --- p.177 / Chapter Chapter 6 --- Reference / Reference --- p.180

Introdução: O câncer da confluência biliopancreática tem alta letalidade e prognóstico reservado, atribuído à associação da agressividade biológica e ao quadro clínico silencioso. A inflamação tem papel fundamental no desenvolvimento e na progressão da caquexia, induzida pela expressão de citocinas produzidas pelo tumor e/ou liberadas pelo sistema imunológico. Nos últimos anos, tem sido documentada a variabilidade na expressão gênica de citocinas que regulam os mecanismos envolvidos na caquexia neoplásica. Objetivos: Em amostras de sangue periférico de pacientes com neoplasia da confluência biliopancreática, avaliar a concentração da interleucina 6 (IL-6), fator de necrose tumoral alfa (TNF-?), interferon-gama (INF- ?) e interleucina 10 (IL-10), além da expressão gênica dessas citocinas, do receptor tipo 1 do fator de necrose tumoral (TNFR1), do receptor tipo 2 do fator de necrose tumoral (TNFR2), da zinco-alfa2-glicoproteina (ZAG) e do receptor ativado por proliferador de peroxissoma gama (PPAR-?). Além disso, o estudo visou identificar possíveis diferenças nos dados avaliados entre os pacientes classificados de acordo com a presença ou não de caquexia neoplásica. Casuística: O estudo transversal foi conduzido com 17 pacientes adultos de ambos os gêneros em pré-operatório imediato de neoplasia da confluência biliopancreática (Grupo Câncer), além de 15 indivíduos controles em pré-operatório de herniorrafia (Grupo Controle). Os pacientes com neoplasia foram classificados de acordo com a presença de caquexia (Subgrupo Caquexia, n=8) e aqueles sem diagnóstico de caquexia (Subgrupo Semcaquexia, n=9). Métodos: A ingestão alimentar e a composição corporal foram avaliadas em todos os voluntários. O questionário de fadiga foi aplicado nos indivíduos com neoplasia. O diagnóstico de caquexia foi feito a partir de critérios pré-estabelecidos. As citocinas inflamatórias IL-6, TNF-?, INF-? e IL-10 foram dosadas no sangue periférico. A expressão gênica dessas citocinas inflamatórias, dos receptores de TNF-?, da ZAG e do PPAR-? foi feita em sangue total. A análise estatística foi realizada com o auxílio de software Statistica, versão 8.0®. Resultados: Não houve diferença na ingestão energética [1827 (1489-2166) vs 1691 (1380-2003) kcal, p=0,56] e proteica [91,6 (74-109) vs 101 (89-114) g, p=0,30] dos indivíduos com câncer ou controles, exceto pela maior ingestão de lipídeos [69,0 (53,5-84,5) vs 42,7 (33,4-52,1) g, p=0,01] e menor consumo de vitamina A [382 (152-612) vs 1346 (1032-1659) ?g, p=0,001] no Grupo Câncer em relação ao Grupo Controle, respectivamente. Houve perda de peso em relação ao habitual em 15 dos 17 pacientes (13,1 ? 11,0%) antes do procedimento cirúrgico, embora as variáveis de composição corporal estivessem semelhantes entre os dois grupos de estudo. Os pacientes com neoplasia apresentavam menores concentrações plasmáticas de albumina [3,8 (3,5-4,0) vs 4,4 (4,3-4,5) g/dL, p<0,001], transferrina [166 (140-192) vs 241 (212-270) mg, p=0,001], ferro [80,8 (67,7-93,9) vs 107 (82-131) ?g/dL, p=0,003], zinco [79,9 (66-93,7) vs 97,4 (88-107) ?g%, p=0,001] e vitamina A [0,7 (0,6-0,8) vs 1,3 (1,0-1,5) umol/L, p<0,001], além de maiores níveis de glicemia [115 (118-193) vs 98,6 (82-115) mg/dL, p=0,003], proteína C reativa [2,7 (0,9-4,5) vs 0,2 (0,2-3,3) mg/dL, p<0,001], ferritina [985 (347-1623) vs 170 (85-255) ng/dL, p=0,003] e cobre [147 (122-177) vs 107 (92-121) ?g%, p=0,03] em relação aos controles, respectivamente. A concentração sérica das citocinas IL-6 [7,2 (4,2-10,1) vs 2,0 (1,4-2,5) pg/mL, p<0,001], TNF- ? [24,6 (18,7-30,5) vs 15,2 (11,3-19,1) pg/mL, p=0,02] e IL-10 [13,3 (8,5-18,2) vs 4,4 (2,8- 6,1) pg/mL, p<0,001] foram maiores no Grupo Câncer. O RNAm do INF-? (p=0,008), 8 TNFR1 (p=0,003), IL-10 (p=0,002) e PPAR-? (p=0,002) foram mais expressos nos pacientes com neoplasia. O Subgrupo Caquexia apresentou menor ingestão energética (p=0,03) e proteica (p=0,04), maior intensidade de fadiga (p=0,003), maior perda de peso (p=0,02) e menores níveis séricos de zinco (p=0,05). Dentre as citocinas, apenas a concentração de IL-6 (p=0,04) foi maior no Subgrupo Caquexia, enquanto que a expressão gênica do INF-? (p=0,04) foi maior nos pacientes com caquexia. Conclusões: Apesar da perda de peso, os marcadores de ingestão alimentar e composição corporal foram pouco alterados nos pacientes com neoplasia da confluência biliopancreática. As alterações laboratoriais foram evidentes nos pacientes com neoplasia, mostrando uma resposta inflamatória sistêmica. O aumento da expressão gênica da IL-10 sugere que as células do sangue periférico estão envolvidas no aumento sérico desta citocina. Apesar do aumento da concentração sérica do IL-6 e TNF-? nos pacientes com neoplasia, não houve aumento da expressão gênica dessas citocinas em sangue periférico. Tais dados sugerem que a IL-6 e o TNF-? são produzidos por outras células do sistema imune, distintas dos macrófagos circulantes. O aumento da expressão gênica de INF-? no sangue periféricos dos pacientes com neoplasia não foi acompanhado por maior concentração sérica dessa citocina, por possíveis mecanismos epigenéticos. Os genes do PPAR-? e do TNFR1 foram mais expressos nos pacientes com neoplasia. A caquexia foi definida em 8 pacientes, que apresentaram maior perda ponderal e menor ingestão nutricional. A concentração sérica de IL-6 foi maior no Subgrupo Caquexia, indicando a relação entre essa citocina e o estado caquético. Embora a concentração sérica de INF-? fosse semelhante entre sujeitos com ou sem caquexia, a expressão gênica dessa citocina foi maior nos pacientes caquéticos. / Introduction: Biliopancreatic confluence cancer has a high lethality and a reserved prognosis, attributed to the association of biological aggressiveness and the silent clinical picture. Inflammation plays a key role in the development and progression of cachexia, induced by the expression of cytokines produced by the tumor and/or released by the immune system. In recent years, it has been documented the variability in the genetics of cytokines regulating the mechanisms involved in neoplastic cachexia. Objectives: To evaluate the concentration of interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-?), interferon-gamma (IFF-?) and interleukin 10 (IL-10) in samples of peripheral blood from patients with biliopancreatic confluence neoplasia, in addition to the gene expression of these cytokines, tumor necrosis factor receptor 1 (TNFR1), type 2 receptor tumor necrosis factor (TNFR2), zinc receptor alpha2-glycoprotein (ZAG) and peroxisome proliferator-activated gamma (PEAR-?). Besides, this study aimed to identify possible differences in the data among patients who were classified according to the presence or absence of neoplastic cachexia. Casuistic: The cross-sectional study was carried out with 17 patients of both genders in the immediate preoperative period of neoplasia of the biliopancreatic confluence (Cancer Group), in addition to 15 individual control in the preoperative period of hernia removal surgery (Control Group). Patients with neoplasia were classified according to the presence of cachexia (Subgroup Cachexia, n=8) and those without it (Subgroup Without-cachexia, n=9). Methods: Food intake and body composition were evaluated in all volunteers. The fatigue questionnaire was applied in individuals with neoplasia. The diagnosis of cachexia was based on pre-established criteria. Inflammatory cytokines IL-6, TNF-?, INF-? and IL-10 were measured in peripheral blood. The gene expression of inflammatory cytokines, TNF-?\' receptors, ZAG and PPAR were made in whole blood. Statistical analysis was performed using Statistica software, version 8.0®. Results: There was no difference in energy intake [1827 (1489-2166) vs 1691 (1380-2003) kcal, p=0.56] and protein [91.6 (74-109) vs 101 (89-114) g, p=0.30] of individuals with cancer or controls, except for the higher lipid intake [69.0 (53.5-84.5) vs 42.7 (33.4-52.1) g, p=0,01] and lower intake of vitamin A [382 (152-612) vs 1346 (1032-1659) ,µg, p=0.001] in the Cancer Group related to Control Group, respectively. There was weight loss from the usual in 15 of the 17 patients (13.1 + 11.0%) prior to the surgical procedure, although body composition variables were similar between the two study groups. Patients with neoplasia had lower plasma albumin concentrations [3.8 (3.5-4.0) vs 4.4 (4.3-4.5) g/dL, p <0.001], transferrin [166 (140-192) vs 241 (212-270) mg/dL, p=0,001], iron [80.8 (67.793.9) vs 107 (82-131) ,µg/dL, p=0.003], zinc [79.9 (66-93.7) vs 97.4 (88-107) µg%, p=0.001] and vitamin A [0.7 (0.6-0.8) vs 1.3 (1, 0-1.5) umol/L, p <0.001], as well as higher glycemia levels [115 (118-193) vs 98.6 (82-115) mg/dL, p=0.003], C-reactive protein 2.7 (0.9-4.5) vs 0.2 (0.2-3.3) mg/dL, p <0.001], ferritin [985 (347-1623) vs 170 (85-255) ng/dL, p=0.003] and copper ~147 (122-177) vs 107 (92-121) µg%, p=0.03] relative to the controls, respectively. The serum concentration of IL-6 cytokines [7.2 (4.2-10.1) vs. 2.0 (1.4-2.5) pg/mL, p <0.001], TNF-? [24.6 (18.7-30.5) vs 15.2 (11.3-19.1) pg/mL, p=0.02] and IL-10 [13.3 (8.5-18.2) vs 4.4 (2.8-6.1) pg/mL, p=0.008) were higher in the Cancer Group. The mRNA of INF-? (p=0.008), TNFR1 (p=0.003), IL-10 (p=0.002) and PPAR-? (p=0.002) were more expressed in patients with neoplasia. The Caquexy Subgroup had lower energetic (p=0.03) and protein intake (p=0.04), higher fatigue intensity (p=0.003), greater weight loss (p=0.02) and lower serum levels of zinc (p=0.05). Among the cytokines, only a concentration of IL-6 (p=0.04) was higher than the Cachexia Subgroup, whereas the gene expression of INF-? (p=0.04) was higher in patients with cachexia. Conclusions: Despite the weight loss, dietary intake markers and body composition were slightly altered in patients with biliopancreatic confluence neoplasia. In patients with neoplasia, laboratory findings were evident showing a systemic inflammatory response. The increasing gene expression of IL-10 suggests that peripheral blood cells are involved in the serum increase of this cytokine. Despite the increased concentration of IL-6 and TNF-? in neoplasia patients, there was no increase in gene expression in peripheral blood cytokines. These data suggest that IL-6 and TNF-? are produced by other cells of the immune system, other than circulating macrophages. The increasing gene expression of INF-? in the peripheral blood of patients with neoplasia was not accompanied by a higher serum concentration of this cytokine, due to possible epigenetic mechanisms. The PPAR-? and TNFR1 genes were more expressed in patients with neoplasia. Cachexia was defined in 8 patients, who presented greater weight loss and lower nutritional intake. The serum concentration of IL-6 was higher in the Cachexia Subgroup, indicating the relation between this cytokine and the cachectic state. Although the serum INF-? concentration was similar between individuals with or without cachexia, the gene expression of this cytokine was higher in cachectic patients