A angiogênese é um processo essencial para a formação placentária. Dentre os muitos fatores envolvidos neste evento biológico, destaca-se de forma preponderante o Fator de Crescimento Endotelial Vascular (VEGF), cuja produção é controlada por genes, que podem apresentar variantes polimórficas diversas. As conseqüências funcionais destas variações não estão ainda completamente elucidadas, embora algumas tenham sido já associadas a um comprometimento da angiogênese em diversos sistemas biológicos. Por outro lado, alterações morfológicas na formação, distribuição e arranjo dos vasos placentários foram descritas na placenta de gestantes com distúrbios hiperglicêmicos como a Diabete mellitus e a hiperglicemia moderada. Particularmente nos casos de hiperglicemia moderada, a adaptação vascular observada parece estar relacionada à manutenção da capacidade funcional placentária. Na diabete, este achado foi correlacionado à expressão protéica e gênica atípica de determinados fatores angiogênicos na interface materno-fetal, enquanto que em outras condições hiperglicêmicas a provável participação destes fatores ainda não foi completamente determinada. Desta forma, partindo da hipótese de que as alterações placentárias vasculares em gestantes hiperglicêmicas podem ser decorrentes da expressão ou produção atípica do VEGF e que esta por sua vez pode ser decorrente de polimorfismos gênicos, o objetivo deste estudo foi avaliar polimorfismos do gene deste fator de crescimento em placentas de gestantes com distúrbios hiperglicêmicos. Foi realizada a análise dos polimorfismos -460C/T, -634G/C e 936C/T do gene do VEGF em fragmentos de vilos coriônicos de gestantes normoglicêmicas, com hiperglicemia moderada e com Diabete mellitus prévia à gestação, denominados respectivamente de grupo IA, IB e IIB. Todo o material foi fornecido pelo Serviço de Diabetes e Gravidez da Faculdade de Medicina de Botucatu UNESP. A genotipagem dos polimorfismos do VEGF foi realizada através da Reação de Polimerase em Cadeia (PCR), seguida de digestão por enzima de restrição. As freqüências de distribuição genotípica e alélica dos polimorfismos do VEGF foram comparadas entre os grupos hiperglicêmicos (IB e IIB) e 6 o grupo controle. As freqüências genotípicas estavam em equilíbrio de Hardy-Weinberg. Foram observadas diferenças significantes nas freqüências genotípicas e alélicas do polimorfismo -460C/T do gene do VEGF em placentas de gestantes hiperglicêmicas (grupos IB e IIB) quando comparadas entre si, mas não com o grupo controle. Diferenças estatísticas nas freqüências genotípicas e alélicas do polimorfismo -634G/C VEGF foram detectadas nos grupos hiperglicêmicos em geral comparados ao grupo controle. Não foram identificadas diferenças nas freqüências genotípicas e alélicas do polimorfismo 936C/T do VEGF entre os grupos avaliados. Em conjunto, nossos achados sugerem que a resposta placentária a distúrbios hiperglicêmicos pode estar correlacionada à presença dos dois polimorfismos: -460C/T e 634C/G. No entanto, nenhuma correlação direta funcional positiva foi observada com a angiogênese observada nas placentas de gestações associadas à hiperglicemia moderada. / The angiogenesis is a process essential for placental formation. Among the many factors involved in this biological event, the factor most active is the vascular endothelial growth factor (VEGF), whose production is controlled by genes, which may present different polymorphic variants. The functional consequences of these changes are not yet fully elucidated, although some have been associated with an impairment of angiogenesis in various biological systems. Moreover, morphological changes in distribution and arrangement of the placental vessels were described in the placenta of pregnant women with hyperglycemic disorders such as diabetes mellitus and mild hyperglycemia. Particularly in cases of mild hyperglycemia, the observed vascular adaptation seems to be related to the maintenance of functional placenta. In diabetes, this finding was correlated with the atypical gene and protein expression of certain angiogenic factors in maternalfetal interface, while in other hyperglycemic conditions the likely participation of these factors has not been fully determined. Thus, based on the hypothesis that placental vascular changes in hyperglycemic pregnant women may be due to atypical expression or production of VEGF and that this in turn may be caused by gene polymorphisms, the objective of this study was to assess polymorphisms of this gene factor growth in placentas of women with hyperglycemic disorders. The analysis of the polymorphisms of the VEGF gene was performed in fragments of chorionic villi of normoglycemic, mild hyperglycemic and diabetic pregnant women, respectively named of group IA, IB and IIB. All specimens were provided by the Serviço de Serviço de Diabetes e Gravidez da Faculdade de Medicina de Botucatu UNESP. Genotyping of the VEGF gene polymorphisms -460C/T, -634G/C and 936C/T was done by the polymerase chain reaction (PCR) and restriction fragment length polymorphism methods. The frequency of VEGF alleles and genotype distribution were compared among the hyperglycemic and control groups. The genotype frequencies were in Hardy-Weinberg equilibrium. We have observed significant differences of VEGF -460C/T polymorphism between the hyperglycemic placentas (IB X IIB), but not compared to control group. Statistical differences in the genotypic and allelic frequencies of C of VEGF -634G/C were detected 8 in hyperglycemic groups in general in comparison to controls. There were no significant differences in allelic or genotype frequencies among the evaluated groups. Taken together our findings suggest that the placental response to hyperglycemic disturbs may be correlated to the presence of both -460C/T and -636C/G polymorphism of the VEGF gene. However, no direct functional correlation was detected between these VEGF polymorphisms and the intense angiogenesis seen in placentas of mild hyperglycemiaassociated gestation.

As doenças cardiovasculares e o diabete melito fazem parte das DCNT prioritárias da OMS, devido às altas taxas de morbimortalidade e incapacidade que geram a cada ano. Estima-se que no mundo existam 387 milhões de diabéticos e outros 316 milhões de pessoas com características de risco, como pré-diabete. Cerca de 60% dos pacientes com DM2 desenvolvem doença cardiovascular, a qual inicia de forma concomitante aos distúrbios do metabolismo da glicose, podendo existir mecanismos fisiopatológicos comuns entre as doenças. Metade dos eventos coronarianos, inclusive a morte súbita, ocorrem em indivíduos assintomáticos, evidenciando a necessidade de novos marcadores precoces, já que em muitos deles a morte é a primeira manifestação. Recentemente, tem sido sugerido que os miRNAs envolvidos na regulação da expressão gênica podem ser caracterizados como biomarcadores em diversas doenças. Nosso objetivo é identificar alterações no perfil de miRNAs circulantes em indivíduos euglicêmicos e pré-diabéticos com e sem aterosclerose subclínica, utilizando a tecnologia de qPCR Arrays, com a finalidade de identificar candidatos a biomarcadores moleculares dessa condição. Encontrou-se que a aterosclerose subclínica esteve associada com o envelhecimento, a menopausa, etnia branca, dislipidemia, resistência à insulina, o aumento da adiposidade, leptina e do TNF-α. O aumento do miR98-5p e a diminuição dos miRNAs miR-212-3p, miR-145-5p, miR-93-5p, miR15a-5p, miR-19a-3p, miR32-5p levaram a ativação da via de sinalização da aterosclerose. Os resultados sugerem que a inflamação foi o principal mecanismo associado com o desenvolvimento de aterosclerose subclínica neste estudo. / Cardiovascular Disease and Diabetes Mellitus are relevant NCDs for the WHO. It´s estimated that, worldwide, there are 387 millions of diabetics and 316 millions of people with risk characteristics like prediabetes. About 60% of patients with T2DM develops cardiovascular disease, which starts at the same time as disorders of glucose metabolism, there may be common pathophysiological mechanisms among diseases. Half of coronary events, including sudden death, occurs in asymptomatic individuals. This fact, highlights the need for new early markers of the disease, especially in asymptomatic patients, since in many of them, death is the first manifestation. It has been recently suggested that miRNAs involved in pos-transcriptional regulation of gene expression, could be characterized as biomarkers of diseases. Our goal is to identify changes in the profile of circulating microRNAs in euglycemic and prediabetic patients with or without subclinical atherosclerosis, by quantitative Polymerase Chain Reaction array (qPCR Array) in order to find molecular biomarkers of this condition. We found that subclinical atherosclerosis was associated with aging, menopause, white ethnicity, dyslipidemia, insulin resistance, increased adiposity, leptin and TNF-α. The up-regulation of miR98-5p and the down-regulation miR-212-3p, miR-145-5p, miR-93-5p, miR15a-5p, miR-19a-3p, miR32-5p led to activation of the signaling pathway of atherosclerosis. The results suggest that inflammation was the main mechanism associated with the development of subclinical atherosclerosis in this study.

Diabete Melito (DM) é uma endocrinopatia frequentemente diagnosticada na clínica de pequenos animais e desenvolve-se pela deficiência relativa ou absoluta de insulina, sendo caracterizada pela hiperglicemia crônica. Complicações associadas ao DM são frequentes, dentre elas podemos citar catarata e retinopatia. A catarata, que é a oftalmopatia mais frequente nos cães, impossibilita à fundoscopia e nestes casos a avaliação da função retiniana pode ser feita com o auxílio do eletrorretinograma de campo total. O objetivo deste estudo foi avaliar a progressão da retinopatia diabética por meio de eletrorretinograma (ERG) de campo total pré e pós-operatória (180 dias) em cães portadores de catarata madura e hipermadura submetidos à facoemulsificação. Vinte e quatro cães (Grupo Diabético (DM) = 12 cães; Grupo Não Diabético (NDM) = 12 cães) de raças diversas foram avaliados. Previamente ao ERG, todos os cães foram submetidos ao exame oftalmológico completo e ultrassom ocular. Todos os cães foram sedados seguindo o mesmo protocolo. ERG de campo total foi realizado com sistema eletrodiagnóstico Veris 2000 e cúpula de estimulação (Ganzfeld), segundo o protocolo da ISCEV contendo 5 respostas. Para obtenção dos registros utilizou-se eletrodo bipolar Burian Allen. Avaliou-se a amplitude pico a pico das 5 respostas e o tempo de culminação da onda-b na resposta de bastonetes, máxima resposta, cones e flicker para cada registro em condições escotópicas e fotópicas. As respostas obtidas em cada olho no momento basal e após 6 meses foram comparadas por meio do teste de Wilcoxon. O teste de Mann-Whitney foi utilizado para comparação de respostas dos olhos operados (OP) com os olhos contralaterais (OC), assim como para comparação entre os grupos DM e NDM. O efeito da cirurgia e do tempo de DM foi ainda avaliado por meio do cálculo das diferenças entre as respostas no momento pré e pós 6 meses de cada olho (delta OP e delta OC). Observou-se diferenças entre OP e OC, no momento pré, nas respostas dos bastonetes e da máxima resposta (amplitude e tempo de culminação onda-b). No momento pós, foram observados diferenças entre OP e OC no tempo de culminação dos bastonetes e flicker, e nas amplitudes do potencial oscilatório, cones e flicker. No grupo NDM, não foram observados diferenças entre os momentos pré e pós assim como entre OP e OC. Quando comparadas as respostas dos grupos DM e NDM, observou-se valores significativamente maiores no grupo DM, no tempo de culminação da onda-b na máxima resposta do OP no momento pós, no tempo de culminação da onda-b na máxima resposta OC no momento pós e no tempo de culminação da onda-b da resposta de cones no momento pré cirurgia. Não foram observados diferenças entre delta OP e delta OC dentro de cada grupo e entre os grupos. Todos os pacientes que participaram do estudo recuperaram a visão após a cirurgia, sugerindo que a inflamação pós cirúrgica foi controlada com a medicação prescrita nos 2 grupos estudados. Conclui-se que a facoemulsificação representa mínimo impacto na evolução da retinopatia diabética em cães. A remoção cirúrgica da catarata não deve ser contraindicada nesta espécie, independentemente se o animal é ou não portador de DM, por resultar em melhoria da visão. / Diabetes mellitus (DM) is a common endocrinopathy in the small animals practice and it develops as a relative or absolute insulin deficiency characterized by a chronic hyperglycemia. There are frequent complications, such as cataracts, retinopathy, resulting in visual impairment. Cataracts are dogs most frequent ophthalmic affection and, since it prevents fundoscopic examination, full-field electroretinography is used for retinal functional evaluation. Therefore, to provide a better understanding of diabetic retinopathy (DR) progression in dogs with mature and hypermature cataracts undergoing phacoemulsification surgery, this study aims to analyze the full-field electroretinogram prior and 180 days post cataracts removal. Twenty four dogs (diabetic group (DM) = 12 dogs and non-diabetic group (NDM) = 12 dogs), from different breeds were evaluated. Before the ERG, all animals were submitted to a complete ophthalmic examination and ocular ultrasonography. All dogs were sedated following the same anesthetic protocol. The full-field ERG was recorded using 2000 VERIS system and a dome stimulator (ganzfeld) according to ISCEV five response standard. The recordings were obtained using bipolar Burian Allen electrodes. Peak to peak amplitude five response and b-wave culmination time in rod responses, maximum response, cone and flicker were evaluated in scotopic and photopic conditions. The responses obtained in each eye at baseline and after 6 months were compared using the Wilcoxon test. The Mann-Whitney test was used to compare the responses of the operated eyes (OP) and the contralateral eyes (OC) as well as for comparison between groups DM and NDMAs. The effect of surgery and duration of DM was assessed by calculating the differences between the responses prior and six months after surgery, of each eye (delta delta OP and OC). There were differences between OP and OC, in the baseline values, in rod responses and maximum response (amplitude values and b-wave implicit time). After phacoemulsification, differences were observed between OP and OC in b-wave implicit time of rods and flicker, and the amplitudes of the oscillatory potentials, cone and flicker. In the NDM group, no differences were observed between pre and post evaluations between OP and OC. Comparing the responses of DM and NDM groups, there were significantly higher values in the DM group, in b-wave implicit time and in maximum response of OP after caratact removal, and also, of the b-wave implicit time in the maximum response in OC after surgery and, finally, in the b-wave implicit time of cone response prior to the surgery. No differences were observed between delta OP and delta OC within each group and between groups. All patients recovered vision after surgery, suggesting that post-surgery inflammation was controlled by the prescribed medication in both groups. The phacoemulsification surgery did not appear to have a great impact on diabetic retinopathy evolution in dogs. Cataract surgery should not be contraindicated in this species, regardless of the presence of DM, as it can result in sight improvement.

O aumento da prevalência do diabete melito do tipo 2 (DM2) é intenso e implica ampla busca pela prevenção e tratamento da doença. Estudos têm mostrado a participação do zinco na síntese, secreção e via de sinalização da insulina e sobre o controle glicêmico. Este trabalho objetivou analisar o mecanismo de ação do zinco no controle da secreção de insulina e no controle glicêmico, de modo a entender se a suplementação com o zinco previne ou retarda a manifestação do DM2. O projeto foi aprovado pela CEUA-ICB (USP). Camundongos machos C57BL/6 foram divididos em 4 grupos experimentais: dieta controle (NFD); dieta controle suplementada com ZnCl2 (NFDZ); dieta hiperlipídica (HFD); e dieta hiperlipídica suplementada com ZnCl2 (HFDZ). A massa corporal, a ingestão de ração e água e a glicemia foram acompanhados semanalmente. Testes intraperitoneais de tolerância à glicose (ipGTT) e à insulina (ipITT) foram realizados na 14ª semana de tratamento. Completado as 15 semanas de tratamento a glicemia, a insulinemia e a zincemia foram analisadas, sendo aplicados os testes de HOMA-IR e HOMA-&#946;. Em ilhotas pancreáticas isoladas foi analisada a secreção estática de insulina em diferentes concentrações de glicose. O teste de captação e metabolismo de glicose foi feito no músculo sóleo e a análise do conteúdo das proteínas AKT e GSK3-&#946; foi feita no músculo sóleo e no fígado. Os dados (média±SEM) foram analisados por Two-way ANOVA com pós-teste Bonferoni ou por teste t de Student (P 0,05). A suplementação com zinco melhorou a disfunção glicêmica induzida por dieta hiperlipídica, sem no entanto afetar a resistência à insulina ou a secreção de insulina por ilhotas isoladas. / The increase in prevalence of type 2 diabetes mellitus (DM2) is intense and implies broad quest for prevention and treatment of disease. Studies have shown the participation of zinc in the synthesis, secretion and signaling pathway of insulin and the glucose control. This study aimed to analyze the mechanism of action of zinc in the control of insulin secretion and glucose control in order to understand whether supplementation with zinc prevents or delays the manifestation of DM2. The project was approved by CEUA-ICB (USP). Male mice C57BL/6 were divided en 4 groups: control diet (NFD); control diet supplemented with ZnCl2 (NFDZ); high fat diet (HFD); and high fat diet supplemented with ZnCl2 (HFDZ). Body weight, feed intake and water and the glucose levels were monitored weekly. Intraperitoneal glucose tolerance test (ipGTT) and insulin (ipITT) were performed at the 14th week of treatment. Completing the 15 weeks of the treatment glycemia, insulinemia and zincemia were analyzed, being applied HOMA-IR and HOMA-&#946; tests. In isolated islets was assessed the static insulin secretion at different glucose concentrations. The uptake and glucose metabolism test was done in the soleus muscle and the content analysis of the AKT and GSK3-&#946; protein was made in the soleus muscle and liver. The data (mean ± SEM) were analyzed by two-way ANOVA with Bonferoni post-test or Student\'s t test (P < 0,05). Zinc supplementation improves glucose dysfunction induced by high fat diet, without nonetheless affecting insulin resistance and insulin secretion by isolated islets.

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.

Diabete melito é umas das principais endocrinopatias, caracterizada pela deficiência relativa ou absoluta de insulina, que pode resultar em diversas manifestações oculares, sendo as mais frequentes a retinopatia diabética e a catarata. Retinopatia diabética (RD) é uma microangiopatia que afeta primeiramente as arteríolas pré-capilares, capilares, vênulas pós-capilares e vasos de maior calibre, causando incompetência funcional e anatômica dos vasos retinianos. A hiperglicemia pode ser a causa mais provável da lesão retiniana, interferindo nas vias de metabolismo celular e no processo de transdução. Os achados fundoscópicos incluem: microaneurismas, dilatação e tortuosidade das vênulas retinianas, hiper-refletividade da área tapetal e exsudatos coriorretinianos. Como a catarata impossibilita a fundoscopia, a prevalência da retinopatia diabética nos cães não esta totalmente esclarecida, sugerindo que esta seja na forma não proliferativa. Objetivou-se neste estudo, determinar a prevalência das alterações fundoscópicas relacionadas à retinopatia diabética em cães, com auxílio de documentação fotográfica (retinografia) e tomografia de coerência óptica (OCT). Vinte e dois cães diabéticos, 18 fêmeas e quatro machos, com idade variando de seis a 15 anos, provenientes do Serviço de Oftalmologia da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, foram submetidos a acompanhamento por meio da retinografia durante o período de 12 meses. Para realização do exame de OCT foram selecionados oito cães dos 22 animais avaliados, quatro fêmeas e quatro machos, com idades variando de seis a 15 anos para compor o grupo diabete melito (GDM), e nove cães sendo cinco fêmeas e quatro machos, com idades entre quatro e 15 anos, sem quaisquer alterações oculares e não diabéticos formaram o grupo controle (GCO). Os cães diabéticos acompanhados durante 12 meses apresentaram alteração vascular, microaneurismas e hemorragias, e alterações na coloração e refletividade da área tapetal. Sendo que dois cães, dos 22 animais avaliados, apresentaram hemorragia em mancha no último período de avaliação, e um cão apresentou focos hemorrágicos durante todo o período de avaliação. A espessura e arquitetura retiniana realizada pela OCT nos cães diabéticos, demonstrou afinamento das camadas da retina e perda da estratificação em comparação com os animais controle (198 &micro;m versus 219 &micro;m, respectivamente), sendo esta redução estatisticamente significante. Com os achados retinográficos deste estudo podemos confirmar que as lesões são compatíveis com a RD não proliferativa sem comprometimento visual, e baseado nas imagens da OCT pode-se sugerir que a diabete melito, no cão, cause neuropatia retiniana como descrito em humanos diabéticos / Diabetes mellitus is one of the most frequent endocrine disorders, characterized by relative or absolute deficiency of insulin, which can induces several ocular manifestations, among them diabetic retinopathy and cataract. Diabetic Retinopathy is a microangiopathy that affects the precapillary, arterioles, capillaries, postcapillary venules, and the large vessels, causing them to be functionally and anatomically incompetent. Hyperglycemia seems to be the most probably cause of retinal damage, interfering in the cellular metabolism process and in transduction process. The fundoscopic findings are microaneurysm, retinal venular dilatation and tortuosity, hyperreflectivity of tapetal area and chorioretinal exsudates. Because the cataract precludes fundoscopic examination, the diagnosis of diabetic retinopathy in dogs is not completely elucidated, but suggests that disease is nonproliferative form. The aim of this study was to determine the prevalence of fundus changes related to diabetic retinopathy in dogs, with photography documentation (fundus camera) and optical coherence tomography (OCT). Twenty-two diabetic dogs, 18 females and four males, from six to 15 years, from Serviço de Oftalmologia da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, underwent monitoring by retinography during the period 12 months. For the OCT examination were selected eight dogs of the 22 evaluated animals, four females and four males, from six to 15 years formed the group diabetes mellitus (GDM), and nine dogs with five females and four males, from 4 to 15 years, with no ocular alterations and non-diabetic formed the control group (GC). Diabetic dogs followed for 12 months showed vascular changes, microaneurisms and hemorrhage, and changes in colour and reflections of tapetal area. Two dogs, among the 22 animals studied, presented hemorrhage stain in the last period, and one dog presented hemorrhage focus throughout the evaluation period. The thickness and retinal architecture performed by OCT in diabetic dogs showed thinning of the retinal layers with loss of stratification compared to control animals (198 &micro;m versus 219 &micro;m, respectively), with a statistically significant difference. Based on fundus findings of this study we can confirm that the lesions are compatible with DR nonproliferative without visual impairment. The OCT images may suggest that diabetes mellitus in the dog causes retinal neuropathy as described in diabetic humans

Diabete melito é umas das principais endocrinopatias, caracterizada pela deficiência relativa ou absoluta de insulina, que pode resultar em diversas manifestações oculares, sendo as mais frequentes a retinopatia diabética e a catarata. Retinopatia diabética (RD) é uma microangiopatia que afeta primeiramente as arteríolas pré-capilares, capilares, vênulas pós-capilares e vasos de maior calibre, causando incompetência funcional e anatômica dos vasos retinianos. A hiperglicemia pode ser a causa mais provável da lesão retiniana, interferindo nas vias de metabolismo celular e no processo de transdução. Os achados fundoscópicos incluem: microaneurismas, dilatação e tortuosidade das vênulas retinianas, hiper-refletividade da área tapetal e exsudatos coriorretinianos. Como a catarata impossibilita a fundoscopia, a prevalência da retinopatia diabética nos cães não esta totalmente esclarecida, sugerindo que esta seja na forma não proliferativa. Objetivou-se neste estudo, determinar a prevalência das alterações fundoscópicas relacionadas à retinopatia diabética em cães, com auxílio de documentação fotográfica (retinografia) e tomografia de coerência óptica (OCT). Vinte e dois cães diabéticos, 18 fêmeas e quatro machos, com idade variando de seis a 15 anos, provenientes do Serviço de Oftalmologia da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, foram submetidos a acompanhamento por meio da retinografia durante o período de 12 meses. Para realização do exame de OCT foram selecionados oito cães dos 22 animais avaliados, quatro fêmeas e quatro machos, com idades variando de seis a 15 anos para compor o grupo diabete melito (GDM), e nove cães sendo cinco fêmeas e quatro machos, com idades entre quatro e 15 anos, sem quaisquer alterações oculares e não diabéticos formaram o grupo controle (GCO). Os cães diabéticos acompanhados durante 12 meses apresentaram alteração vascular, microaneurismas e hemorragias, e alterações na coloração e refletividade da área tapetal. Sendo que dois cães, dos 22 animais avaliados, apresentaram hemorragia em mancha no último período de avaliação, e um cão apresentou focos hemorrágicos durante todo o período de avaliação. A espessura e arquitetura retiniana realizada pela OCT nos cães diabéticos, demonstrou afinamento das camadas da retina e perda da estratificação em comparação com os animais controle (198 &micro;m versus 219 &micro;m, respectivamente), sendo esta redução estatisticamente significante. Com os achados retinográficos deste estudo podemos confirmar que as lesões são compatíveis com a RD não proliferativa sem comprometimento visual, e baseado nas imagens da OCT pode-se sugerir que a diabete melito, no cão, cause neuropatia retiniana como descrito em humanos diabéticos / Diabetes mellitus is one of the most frequent endocrine disorders, characterized by relative or absolute deficiency of insulin, which can induces several ocular manifestations, among them diabetic retinopathy and cataract. Diabetic Retinopathy is a microangiopathy that affects the precapillary, arterioles, capillaries, postcapillary venules, and the large vessels, causing them to be functionally and anatomically incompetent. Hyperglycemia seems to be the most probably cause of retinal damage, interfering in the cellular metabolism process and in transduction process. The fundoscopic findings are microaneurysm, retinal venular dilatation and tortuosity, hyperreflectivity of tapetal area and chorioretinal exsudates. Because the cataract precludes fundoscopic examination, the diagnosis of diabetic retinopathy in dogs is not completely elucidated, but suggests that disease is nonproliferative form. The aim of this study was to determine the prevalence of fundus changes related to diabetic retinopathy in dogs, with photography documentation (fundus camera) and optical coherence tomography (OCT). Twenty-two diabetic dogs, 18 females and four males, from six to 15 years, from Serviço de Oftalmologia da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, underwent monitoring by retinography during the period 12 months. For the OCT examination were selected eight dogs of the 22 evaluated animals, four females and four males, from six to 15 years formed the group diabetes mellitus (GDM), and nine dogs with five females and four males, from 4 to 15 years, with no ocular alterations and non-diabetic formed the control group (GC). Diabetic dogs followed for 12 months showed vascular changes, microaneurisms and hemorrhage, and changes in colour and reflections of tapetal area. Two dogs, among the 22 animals studied, presented hemorrhage stain in the last period, and one dog presented hemorrhage focus throughout the evaluation period. The thickness and retinal architecture performed by OCT in diabetic dogs showed thinning of the retinal layers with loss of stratification compared to control animals (198 &micro;m versus 219 &micro;m, respectively), with a statistically significant difference. Based on fundus findings of this study we can confirm that the lesions are compatible with DR nonproliferative without visual impairment. The OCT images may suggest that diabetes mellitus in the dog causes retinal neuropathy as described in diabetic humans

Le présent travail rend compte des études phytochimiques et biologiques d'extraits non volatils de quatre plantes utilisées au Bénin dans le traitement du diabète. La première partie passe en revue la bibliographie sur les plantes sujettes à notre étude. Dans cette partie, la systématique, l'importance en pharmacopée ainsi que les travaux déjà effectués sur ces plantes ont été présentés. La deuxième partie présente le mode d'extraction et les études chimiques des extraits et les résultats obtenus. La spectrophotométrie a permis de déterminer quelques grandes familles de composés présents dans les extraits : les polyphénols totaux, les flavonoïdes et les tanins tandis que la GC/MS et la LC/MS ont servi à mettre en exergue la présence de composés volatils et non volatils. La troisième partie décrit les tests biologiques in vitro et ex vitro effectués sur les extraits. Les extraits ont montré de manière générale des activités : inhibitrice de l' α-glucosidase, antioxydantes (DPPH, FRAP, ORAC), antimicrobiennes et l'une (Bridelia ferruginea) une activité cytotoxique sur les cellules cancéreuses (PA1, MCF7, PC3, DU-145), avec une efficacité variable d'une plante à une autre. La quatrième partie discute de manière générale des résultats issus des études phytochimiques et des tests biologiques. Parmi les quatre échantillons de plantes sélectionnées pour notre étude, seul l'extrait semi-éthanolique des racines de Ceiba pentandra a une faible teneur en familles de composés dosés et présente des activités biologiques (ci-dessus citées) faibles comparativement aux extraits de Bridelia ferruginea, de Pseudocedrela kotschyi et de Polygonum senegalensis. L'ensemble des résultats tant sur le plan chimique que biologique met en évidence les potentialités des extraits de plantes étudiées, pour une exploitation à des fins thérapeutiquesfutures. / The present work had reported on the phytochemical and biological studies of non-volatile extracts of four plants used in Benin for diabetes treatment. The first part reviewed the bibliography of investigated plants in our study. In this part, the systematic, the importance in the pharmacopoeia and the previous works done on these plants were presented. The second part has presented the extraction method and chemical studies of the extracts and results obtained. The spectrophotometry has permitted to identify some important families of compounds in the extracts: the total polyphenols, flavonoids and tannins whereas the GC / MS and LC/MS were used to highlight the presence of volatile and non-volatile compounds. The third part described the biological tests in vitro and ex vitro carried out on the extracts. The extracts showed in general activities: α-glucosidase inhibition, antioxidant (DPPH, FRAP, ORAC), antimicrobial, and one of them (Bridelia ferruginea) were cytotoxic on cancer cells (PA1, MCF7, PC3, DU-145), with a variable efficiency from one plant to another. The fourth part had discussed in general about the results obtained from phytochemical studies and biological tests. Among the four plants samples selected for our study, only the semi-alcoholic extract of Ceiba pentandra roots had a low-dosed compounds families and presented of this biological activities (cited below) low comparatively to Bridelia ferruginea, Pseudocedrela kotschyi and Polygonum senegalensis extracts. Both of the chemical and biological results highlight the potential of certain species for future exploitation of their non-volatile extract for therapeutic purposes.

Dans ce travail, nous démontrons que l’expression ectopique de Pax4 dans les cellules glucagon+ adultes induit, indépendamment de l’âge, leur néogenèse et transformation en cellules «bêta-like», ce qui entraîne une hypertrophie des îlots et une néogenèse inattendue des îlots. Par l’utilisation de plusieurs approches de traçage, nous démontrons que la conversion des cellules alpha en cellules «bêta-like» médiée par l’expression de Pax4, induit également la mobilisation de précurseurs situés dans ou à proximité des canaux pancréatiques. Ces cellules ré-expriment le gène développemental Ngn3 et adoptent successivement une identité de cellules glucagon+ puis de cellules «bêta-like», suggérant le réveil des mécanismes embryonnaires. Il et à noter que ces processus sont capables de régénérer la totalité de la masse de cellules bêta après plusieurs séries d’induction chimique du diabète. Ces résultats offrent ainsi des perspectives prometteuses pour concevoir de nouvelles stratégies thérapeutiques et régénératrices dans le contexte du diabète du type I. Dans un deuxième chapitre, ce travail décrit nos résultats d'analyse par puce à ADN de pancréas transgénique d’animaux exprimant conditionnellement le gène Pax4 dans les cellules alpha adultes. Cette approche nous permis d'identifier de potentiels gènes cibles de Pax4, qui pourraient jouer un rôle important dans les processus de régénération de la masse de cellules bêta. L’analyse de la fonction de l’un de ces gènes, le facteur de croissance indépendante 1 (Gfi1) est décrite. / In this work we demonstrate that the inducible misexpression of Pax4 in glucagon+ cells age-independently provokes their conversion into beta-like cells and their glucagon shortage-mediated replacement, this process resulting in islet hypertrophy and in an unexpected islet neogenesis. Taking advantage of several lineage-tracing approaches, we show that, upon Pax4-mediated alpha-to-beta-like cell conversion, pancreatic duct-lining precursor cells are mobilized, re-express the developmental gene Ngn3, and successively adopt a glucagon+ and a beta-like cell identity through a mechanism involving the reawakening of the epithelial-to-mesenchymal transition (EMT). It is worth mentioning, that these processes can repeatedly regenerate the entire beta-cell mass, and thereby reverse several rounds of streptozotocin-mediated chemically-induced Type I diabetes. This approach thereby provides promising perspectives to design novel therapeutic regenerative strategies. Aiming to gain further insight into the molecular mechanisms underlying these regeneration and reprogramming processes, and thereby identify new putative targets of interest, a thorough micro array analysis was performed using pancreata from transgenic mice conditionally misexpressing Pax4 in adult alpha-cells. We thereby identified several promising candidate genes, whose gene expression was significantly altered in induces animals. Among these was Growth factor independent 1 (Gfi1): its expression pattern and putative function in the murine pancreas will be described in this work.

Diabetes mellitus is a severe chronic life-long disease. The condition itself introduces a need for patient's lifestyle adjustment to the disease and a number of everyday therapeutic and diagnostic restrictions. Therefore, mental disorders are more common in diabetic patients than in the rest of the population. Biochemical and hormonal connections between mental disorders and diabetes mellitus represent another reason for their higher incidence in diabetic patients. Comorbid mental diseases can further negatively influence the course of diabetes. They are especially depression, anxiety disorders, eating disorders and cognitive disorders including dementia. Type 2 diabetes is also more common in patients with primary mental disease, as is e.g. schizophrenia and bipolar affective disorder. Regarding therapy, psychoactive drugs are used in diabetic patients. It is important to respect the specifics of the underlining disease during drug selection. The main factor for the selection of the medication is, apart of mental problems, the influence on body weight and blood glucose. Mental disorders can be also treated by psychotherapy and psychoeducation. Studies performed in diabetic patients with mental problems suggest the need for intervention in this area. The practical part of the work introduces a...