Introduction: Strength training has been consistently demonstrated in studies as responsible for significant increases in lean mass and metabolic rate, accompanied by significant reductions in body fat weight, using strategies to accelerate this process androgenic anabolic steroids has been enough used by practitioners of this modality. Objective: To evaluate the strength training acting in conjunction with Androgenic Anabolic Steroids on the percentage of body fat of eutrophic rats. Methods: Twentyfour female rats randomly distributed in four groups were used: 1) Sedentary Control (CS) 2) Trained Control (CT) 3) Sedentary Nandrolone Decanoate (DS) 4) Trained Nandrolone Decanoate (DT). Strength training was performed in a squatting apparatus composed of four sets of 12 repetitions, with intensity of 70% of 1RM for eight weeks. On alternate days, the DS and DT groups received daily 5 mg/kg nandrolone decanoate intraperitoneally and the CS and CT groups received only saline solution (0.9%). The data represent the mean ± standard error of the mean. Student's t-test was used for analysis between groups, ns = no statistical difference. Results: After eight weeks of training, the weight between the CS and CT groups were different when compared to the DS and DT groups (there was no statistical difference between groups (CS vs CT, DS vs DT), in this sense, the CT group Who underwent strength training had a 10.8% and 11.2% increase in strength, in the 6th and 8th weeks, respectively, when compared to the CS group. The fatty contents of different territories were evaluated as follows: subcutaneous (SUB), retroperitoneal (RETRO) and periovarian (PERI) fats, which did not identify statistical differences between the groups evaluated. Conclusion: The use of Nandrolone Decanoate in trained rats did not cause changes in weight, strength and adipose tissue. / Introdução: Treinamentos de força têm sido consistentemente demonstrado em estudos como responsáveis por aumentos significativos na massa magra e da taxa metabólica, acompanhada por reduções significativas no peso de gordura corporal, utilizando-se de estratégias para acelerar este processos os esteroides anabólicos androgênicos tem sido bastante utilizados por praticantes desta modalidade. Objetivo: Avaliar o treinamento de força atuando de forma conjunta com Esteroides Anabólicos Androgênicos sobre o percentual de gordura corporal de ratas eutróficas. Métodos: Foram utilizados 24 ratas fêmeas distribuídas randomicamente em quatro grupos: 1) Controle Sedentário (CS) 2) Controle Treinado (CT) 3) Decanoato de Nandrolona Sedentário (DS) 4) Decanoato de Nandrolona Treinado (DT). O treinamento de força foi realizado em aparelho de agachamento composto por quatro séries de 12 repetições, com intensidade de 70% de 1RM durante oito semanas. Em dias alternados, os grupos DS e DT recebiam diariamente Decanoato de nandrolona intraperitoneal 5mg/kg por secção e os grupos CS e CT recebiam somente solução salina (0,9%). Os dados representam a média ± erro padrão da média. Utilizou-se o teste t de Student para análise entre os grupos, ns = sem diferença estatística. Resultados: Após oito semanas de treinamento, o peso entre os grupos CS e CT foram diferentes quando comparados com os grupos DS e DT (não foi observado diferença estatística intergrupos (CS vs CT; DS vs DT)), neste sentido, o grupo CT que foram submetidos ao treinamento de força apresentaram um incremento da força de 10,8% e 11,2%, nas 6ª e 8ª semanas, respectivamente, quando comparado ao grupo CS. Foram avaliados os conteúdos gordurosos de diferentes territórios conforme segue: gorduras subcutâneas (SUB), retroperitoneal (RETRO) e periovariana (PERI) o qual não identificamos diferenças estatísticas entre os grupos avaliados. Conclusão: O uso de Decanoato de Nandrolona nas ratas treinadas não causou alteração, no peso, na força e no tecido adiposo.

A Síndrome do Intestino Curto (SIC) é uma doença complexa que ocorre após extensa ressecção do intestino delgado, levando a uma má absorção de nutrientes e fluidos, uma condição que pode causar diarreia, desnutrição e perda de peso graves com alto risco para o desenvolvimento da osteoporose. Estudos recentes mostram existir ampla interação fisiológica do esqueleto com os diversos sistemas, incluindo o metabolismo energético e o trato digestório. Peptídeos originados não só no tecido adiposo, mas também no intestino como as incretinas [GIP (polipeptídeo trópico insulínico dependente de glicose) e GLP1 (peptídeo 1 tipo glucagon)] modulam a atividade de remodelação óssea. O objetivo principal do atual estudo foi avaliar a relação entre os tecidos adiposos subcutâneo (TAS), visceral (TAV), lipídeos intra-hepáticos (LIH), tecido adiposo da medula óssea (TAMO), bem como do GIP, GLP1, e grelina com a densidade mineral óssea (DMO) em pacientes com SIC. Tratase de um estudo observacional prospectivo composto por dois grupos experimentais pareados por altura, idade e sexo: a) o grupo controle (GC) (n = 18; 9M,9F) e b) o grupo de pacientes com SIC, o qual foi avaliado em 2 ocasiões, com intervalo de um ano entre as análises, sendo denominados SIC0 (n = 14; 7M,7F) e SIC1 (n = 11; 6M,5F). Em comparação com o GC, pacientes com SIC ao longo do estudo apresentaram menor DMO e maior LIH e GIP (p< 0,05). Os valores de TAMO, GLP1 e grelina foram similares entre os grupos. O TAMO teve correlação negativa e significativa com DMO de L3 no GC (r= -0,6; p< 0,05), porém, no grupo SIC esta correlação foi positiva, mas sem significância estatística ao longo do estudo: SIC0 (r= 0,45; p= 0,13) e SIC1 (r= 0,45; p= 0,17). LIH associou-se negativamente com DMO do colo do fêmur (R²= 0,16; p< 0,05) e quadril total (R²= 0,27; p< 0,05). Existe alta prevalência de osteoporose em pacientes com SIC. No entanto, não se observou nem expansão de TAMO e nem relação negativa da DMO com o TAMO. O acesso a calorias parece afetar positivamente a relação entre TAMO e massa óssea. A deposição hepática de lipídeos parece afetar negativamente a massa óssea de pacientes com SIC. / Short bowel syndrome (SBS) is a complex disease that occurs after extensive resection of the small intestine leading to malabsorption of nutrients and fluids, a condition that can cause severe watery diarrhea, dehydration and acute weight loss, developing high risk for the appearance of osteometabolic disease. Studies have shown the progress on the physiological interaction of the skeleton with the various systems, including energetic metabolism and the gastrointestinal tract. Peptides originated not only in adipose tissue but also in the intestine such as incretin [GIP (Glucose-dependent insulinotropic polypeptide) and GLP1 (glucagonlike peptide 1) modulate bone remodeling activity. The main objective of the current study was to evaluate the influence of subcutaneous (SAT), visceral (VAT) adipose tissue, intrahepatic lipids (IHL), bone marrow fat adipose tissue (MAT), as well as the influence of GIP, GLP1, and ghrelin on the bone mineral density (BMD) of SBS patients. It is a prospective observational study composed by two experimental groups matched by height, age and sex: a) the control group (CG) (n = 18; 9M,9F) and b) the SBS group which were evaluated in two occasions with a period between analyzes of one year: named SBS0 (n = 14; 7M,7F) and SBS1 (n = 11; 6M,5F). Compared to CG, SBS patients throughout the study had significantly lower BMD and elevated IHL and GIP (p< 0.05). Values of MAT, GLP1 and ghrelin were similar between groups. MAT was negatively and significantly correlated with L3 BMD in the CG (r = -0.6; p< 0.05) and positively correlated, but not significant with L3 BMD in the SBS group throughout the study: SBS0 (r= 0.45; p= 0.13) and SBS1 (r= 0.45; p= 0.17). IHL was negatively and significantly associated with femoral neck BMD (R²= 0.16; p< 0.05) and total hip BMD (R²= 0.27; p< 0.05). The occurrence of osteoporosis is frequent in SBS patients, but MAT is not increased in these patients and had positive correlation with BMD, although not significant. Access to calories seems to positively affect the relationship between MAT and bone mass. IHL appear to negatively affect bone mass in SBS patients.

INTRODUÇÃO: A obesidade mórbida é um problema de saúde pública. O aumento da massa gorda contribui para a perda de massa livre de gordura e mudanças na força muscular e resistência dos músculos dos membros inferiores (flexores e extensores), que são responsáveis pela independência, mobilidade e capacidade para realizar com segurança as atividades diárias. OBJETIVO: Avaliar a correlação entre a força muscular e composição corporal total e segmentar de acordo com o grau de obesidade. MÉTODO: Foram incluídos no estudo 132 pacientes com obesidade mórbida de ambos os sexos, com idade entre 18 e 60 anos, sendo divididos entre grupo obeso (>= 40Kg/m2 e < 50Kg/m²) e superobeso (>= 50Kg/m2 e < 60Kg/m²). Todos os pacientes realizaram avaliação da composição corporal (bioimpedância elétrica) e da força muscular máxima dos membros inferiores (dinamometria isocinética). RESULTADOS: Não houve diferença significativa entre o valor médio da força muscular absoluta de extensão (156,4 ± 45 Nm vs. 156,4 ± 41 Nm) e flexão (71,5 ± 22 Nm vs. 72,8 ± 22 Nm) entre o grupo obeso e superobeso. O grupo superobeso apresentou redução da força muscular de extensão e flexão corrigida pelo peso corporal e pelo peso dos membros inferiores em relação ao grupo obeso (P < 0,05). A correlação da força muscular com o peso corporal foi fraca (r = 0,37-0,57, P < 0,001) a moderada, enquanto que observou se correlação moderada em relação ao peso dos membros inferiores (r = 0,46-0,61, p < 0,001). CONCLUSÃO: O grupo superobeso apresentou um valor médio na força absoluta de extensão e de flexão. A força muscular diminuída na obesidade grave está diretamente relacionada com a composição corporal dos membros inferiores / BACKGROUND: Morbid obesity is a public health problem. The increase in fat mass contributes to loss of fat free mass and changes in strength and endurance of lower limbs muscles (flexors and extensors) that are responsible for independence, mobility and ability to perform daily activities safely. OBJECTIVE: to evaluate the correlation between the muscle strength and total and segmental body composition according to the obesity grade. METHODS: 132 morbidly obese patients were included in the study of both sexes, aged 18 and 60 years, divided between obese group (>= 40Kg/m2 e < 50Kg/m²) and super obese (>= 50Kg/m2 e < 60Kg/m²). All patients performed a body composition evaluation (bioelectrical impedance analysis) and maximum muscle strength of the lower limbs (isokinetic dynamometry). RESULTS: There was no significant difference between absolute extension (156.4 ± 45 Nm vs. 156.4 ± 41 Nm) and flexion muscle strength (71.5 ± 22 Nm vs. 72.8 ± 22) between the obese and the super obese group. The super obese group presented a reduced extension and flexion muscle strength corrected to body weight and to weight of the lower limbs in compared to obese group (P < 0.05). The correlation of muscle strength relative to body weight was weak to moderate (r = 0.37 - 0.57, P<0.001) whereas strength relative to weight of the lower limbs presented a moderate correlation (r = 0.46 - 0.61, P < 0.001).CONCLUSIONS: Super obese group had a mean value in absolute extension and flexion muscle strength. The decreased muscular strength in severe obesity is directly related to body composition of the lower limbs

INTRODUÇÃO: As células-tronco hematopoiéticas (CTH) do sangue do cordão umbilical (SCU) têm sido utilizadas com sucesso para o tratamento de doenças malignas e não malignas. No entanto, algumas unidades de SCU podem apresentar baixa quantidade de células nucleadas totais (CNT). Algumas abordagens têm sido sugeridas para evitar problemas em relação à baixa concentração de CTH no transplante, como a administração de duas unidades de SCU para o paciente e a expansão ex vivo de CTH. OBJETIVO: Avaliar as taxas de proliferação celular na expansão ex vivo do SCU em sistema de cocultura com células-tronco mesenquimais (CTM) obtidos a partir de diferentes fontes com alta e baixa confluência e adicionando-se ou não coquetel de citocinas no meio de cultura. MÉTODOS: Este estudo foi aprovado pelo Comitê de Ética de Pesquisa (CAPPesq) do Hospital das Clínicas da Faculdade de Medicina da USP. A coleta do SCU (n =10) foi realizada após o nascimento do bebê e expulsão da placenta. O processamento foi realizado utilizando o método de redução de volume, o qual consiste em depleção de eritrócitos. As amostras de CTM provenientes do endotélio vascular do cordão umbilical foram obtidas de doadores diferentes (n=3) e o tecido adiposo (n=3) do inventário do LIM-31. A expansão das CNT e das células com expressão de marcadores CD133+/CD34+ foram observados depois de sete dias de cultura. Além disso, o ensaio para análise de unidades de formadoras de colônias (UFC) foi realizado em todas as amostras antes e depois da expansão do SCU. Para a expansão em sistema de cocultura foi separado dois grupos para ambas as fontes de CTM (Grupo I - cocultura com adição de coquetel de citocinas vs. Grupo II - cocultura sem citocinas). RESULTADOS: Após sete dias, no grupo I com cocultura confluente, a taxa de proliferação de CNT foi duas vezes maior ao comparar com cocultura subconfluente (35 vs. 16 vezes). No mesmo grupo também foi possível evidenciar elevada taxa de proliferação de células CD133+/CD34+. O índice de proliferação das UFC no grupo I aumentou até oito vezes. A cocultura subconfluente tanto do endotélio vascular do cordão umbilical como do tecido adiposo apresentou menor rendimento em comparação as CTM confluentes. A expansão das células na presença de citocinas apresentou maior proliferação celular ao comparar às coculturas sem adição de citocinas. CONCLUSÃO: Este estudo mostrou que para alto rendimento de células do SCU, o sistema de cocultura requer adição de coquetel de citocinas e CTM confluente independentemente da fonte utilizada / INTRODUCTION: Umbilical cord blood (UCB) hematopoietic stem cells have been successfully used for the treatment of both malignant and non-malignant diseases. Nevertheless, some UCB units could have low total nucleated cells (TNC) dose. Several approaches have been suggested to avoid inadequacy problems of hematopoietic stem cells (HSC) number for transplantation, such as administration of two UCB units to the patient and HSC ex vivo expansion. OBJECTIVE: Evaluate UCB ex vivo expansion proliferative rates in a high and low mesenchymal stem cells (MSC) confluence feeder layer obtained from different MSC sources and by adding or not cytokines cocktail into the medium. METHODS: This study was approved by the Research Ethic Committee (CAPPESQ) of Hospital das Clínicas da Faculdade de Medicina da USP. The collection of UCB (n=10) was made after delivery of the infant and the expulsion of placenta. Processing was performed using volume reduction method which consists in red blood depletion. MSC samples from umbilical cord endothelium were obtained from three different donors and adipose tissue (n=3) obtained from LIM31\'s pattern inventory. The total nucleated cell (TNC), expression of hematopoietic surface markers such as CD133+/CD34+ were observed after seven days of culture. Beyond that, colony forming unit assay (CFU) was performed before and after UCB expansion. The expansion by coculture method was observed in two groups (Group I - coculture with cytokines cocktail added vs. Group II- coculture without cytokines cocktail) for both MSCs sources. RESULTS: After seven days, analysis of confluent coculture showed that TNC proliferation rate ware almost 2 times higher than in subconfluent coculture (35 vs. 16-fold) in Group I and also revealed higher proliferative rate in CD133+/CD34+ cells considering. CFU showed similar increase after seven days of culture in comparison of day 0 (up to 8-fold). Subconfluent coculture for both umbilical cord endothelium and adipose tissue showed lower yield compared with those with high MSC confluence. The expansion in the presence of cytokines showed higher cell proliferation compared to the cocultures without addition of cytokines. CONCLUSION: This study showed that coculture system may require the addition of cytokines cocktail in the media and confluent MSC regardless of source for high yield of UCB cells

O consumo de ácidos graxos n-3 tem sido associado à proteção contra a obesidade, inflamação e resistência à insulina. Os n-3 são ligantes fracos dos receptores nucleares PPAR&gamma;, e pela ativação deste podem exercer suas ações metabólicas e anti-inflamatórias. No presente trabalho, foi investigado se o aumento da disponibilidade dos n-3 geneticamente ou por dieta, via ativação de PPAR&gamma;, protege camundongos do desenvolvimento da obesidade, intolerância a glicose e inflamação do tecido adiposo. Foi visto em um modelo com camundongos fat-1 (elevados níveis endógenos de n-3) que dentre as ações dos ácidos graxos n-3, a proteção contra o aumento de peso/adiposidade associada à obesidade, bem como a melhora da intolerância à glicose são dependentes de PPAR&gamma;. Além disso, por meio da utilização de camundongos com deleção de PPAR&gamma; em hepatócitos os dados desse trabalho mostram que os PPAR&gamma; são, ao menos em parte, essenciais ao aumento da oxidação de ácidos graxos induzida pelos n-3 devido à modulação da expressão gênica e protéica de enzimas mitocondriais e peroxissomais. / The intake of n-3 fatty acids have been associated to the protection against obesity, inflammation and insulin resistance. The n-3 fatty acids are ligands of the nuclear receptor PPAR&gamma;, and by the activation of this receptor can promote their metabolic and anti-inflammatory effects. Herein, we investigated whether increasing body n-3 fatty acids levels either genetically or by a n-3 enriched diet protects mice from diet-induced obesity, glucose intolerance and adipose tissue inflammation through PPAR&gamma; activation. Fat-1 mice were protected from diet-induced obesity, glucose intolerance and adipose tissue inflammation. To better investigate PPAR&gamma; involvement in n-3 beneficial actions, mice with genetic deletion of PPAR&gamma; specifically in hepatocytes. In spite of the absence of changes in body weight and glucose homeostasis PPAR&gamma; deletion in hepatocytes completely abolished the increase in liver fatty acid oxidation and hepatic gene expression of genes associated to mitochondrial and peroxisomal activity.

Estudos com células tronco mesenquimais (CTm) têm despertado grande interesse devido a seu promissor potencial terapêutico e representam uma alternativa para o tratamento de diversas patologias em diferentes órgãos, inclusive em transplante renal. A rejeição crônica é um dos maiores desafios no transplante tardio e se caracteriza por perda progressiva da função renal causado pela intensa fibrogênese no aloenxerto. Os tratamentos convencionais com imunossupressores, apesar de reduzirem significativamente as crises de rejeição aguda, não interferem na sobrevida do enxerto a longo prazo. A compreensão dos processos fisiopatológicos da doença depende de seu estudo em modelos experimentais, que são de grande importância pois também propiciam uma melhor compreensão dos possíveis tratamentos. O presente estudo teve como objetivo analisar a terapia com células-tronco mesenquimais derivadas de tecido adiposo (CTmTA) no modelo experimental de transplante renal em ratos, para estudar seu efeito na rejeição crônica e avaliar seu potencial efeito imunomodulador. O modelo foi estabelecido com ratos das linhagens isogênicas Fisher (doador) e Lewis (receptor) e os animais transplantados foram divididos em três grupos: ISO (transplante isogênico de Lewis para Lewis, n=6), ALO (transplante alogênico de Fisher para Lewis, n=6) e ALO+CTmTA (transplante alogênico, tratado com CTmTA, n=6). As CTmTA foram caracterizadas por aderência ao plástico, diferenciação nas linhagens adipogênica, condrogênicas e osteogênicas e por citometria de fluxo. Foram inoculadas 1 x 106 células na região subcapsular renal no dia da realização da nefrectomia unilateral direita (10 dias pós-transplante). Após 6 meses foram realizadas análises dos parâmetros clínicos e laboratoriais, além de análise histológica, imunohistoquímica e PCR em tempo real. As CTmTA foram eficientes em prevenir significativamente a elevação da ureia e da creatinina séricas, manter clearence de creatinina em níveis normais, e prevenir a elevação da fração de excreção de Na+ e K+. Além disso, impediram o desenvolvimento de proteinúria e da hipertensão arterial. A análise histológica mostrou uma redução significativa do infiltrado inflamatório de macrófagos e linfócitos T, além de uma diminuição da fibrose intersticial no grupo ALO+CTmTA. O tratamento com CTmTA reduziu significativamente a expressão relativa dos fatores e citocinas pró-inflamatórios tais como INF-y, TNF-alfa, IL1beta e IL-6, além de aumento importante na expressão de IL-4 e IL-10, conhecidas por seu potencial antiinflamatório. Em conclusão, o tratamento com ADMSC em um modelo experimental de transplante renal pode trazer uma nova abordagem terapêutica para controle da rejeição crônica do enxerto. A aparente modulação da resposta imune observada neste trabalho, pode estar associada a uma possível polarização de macrófagos e células T. Outros estudos pré-clínicos e clínicos são necessários para confirmar nossos resultados / Studies involving mesenchymal stem cells (MSCs) have aroused great interest due to their promising therapeutic potential representing an alternative for the treatment of several pathologies in different organs, including renal transplantation. Chronic rejection is one of the major challenges in late transplantation and is characterized by progressive loss of renal function caused by intense fibrogenesis in the allograft. Conventional immunosuppressive treatments, while significantly reducing acute rejection crises, do not interfere with long-term graft survival. Animal model of kidney transplantation can provide a better understanding of the pathophysiological processes and bring a new path to treat chronic rejection. The aim of this project was to analyze the therapy with mesenchymal stem cells derived from adipose tissue (ADMSCs) in the experimental model of kidney transplantation in rats, focus on chronic rejection and evaluate its potential immunomodulatory effect. The model was established with rats of isogenic strains Fisher (donor) and Lewis (recipient), and the transplanted animals were divided into three groups: ISO (isogenic transplantation from Lewis to Lewis, n = 6), ALO (allogenic transplant from Fisher to Lewis, n = 6) and ALO + ADMSCs (allogenic transplantation, treated with ADMSCs, n = 6). ADMSCs were characterized by adhesion to plastic, differentiation in adipogenic, condrogenic and osteogenic lines and by flow cytometry. One million of cells were inoculated under the renal capsule on the day of the right unilateral nephrectomy (10 days after transplantation). After 6 months, clinical and laboratory parameters were analyzed, as well as histological analysis, immunohistochemistry and real-time PCR. ADMSCs were effective in preventing elevation of serum urea and creatinine, elevation of the Na + and K + excretion fraction as well as maintained creatinine clearence at normal levels. Furthermore, the treatment also prevented the development of proteinuria and preserved blood pressure. Histological analysis showed a significant reduction of macrophages and T cells infiltrate, associated to a decreased of interstitial fibrosis in the ALO + ADMSCs group. In the presence of ADMSCs, there was a significant decrease in the relative expression of INF-y, TNF-alpha, IL1beta and IL-6 factors and pro-inflammatory cytokines, as well as a significant increase in the relative expression of anti-inflammatory cytokines as IL-4 and IL-10. In conclusion, treatment with ADMSC in a transplantation model could open a new approach to control chronic rejection. This apparent modulation of the immune response may be associated with a possible polarization of macrophages and T cells. Further pre-clinical and clinical studies are needed to confirm our findings

Diabetes mellitus tipo 2 (DM2) está associado a um estado de inflamação crônica e ativação do estresse do retículo endoplasmático (ERE). Nesse contexto, são necessários estudos para encontrar alternativas que melhorem o quadro inflamatório, como os ácidos graxos poli-insaturados ômega 3 (n-3 PUFA), um conhecido agente anti-inflamatório. Esse estudo teve por objetivo avaliar o efeito da suplementação do ácido alfa-linolênico (ALA, um n-3 PUFA) no estresse do retículo endoplasmático e no estado inflamatório no tecido adiposo subcutâneo abdominal (TASC) em pacientes com DM2. Foi conduzido um estudo duplo-cego, prospectivo, placebo-controlado. Vinte pacientes com DM2 foram randomizados para suplementação com 3g/dia de ALA ou placebo durante 60 dias. O tecido adiposo foi coletado através de punção aspirativa por agulha fina do abdome antes e após a suplementação e os genes e proteínas foram avaliados através de PCR em tempo real e western blot. Foi encontrada, após suplementação, uma redução da expressão gênica do XBP1 (20%), sXBP1 (70%) e aumento da expressão gênica do GRP78 (150%), confirmado na expressão proteica. Além disso, foi encontrado aumento da expressão gênica da adiponectina (90%) e redução da expressão gênica do IL-6 (80%) e IRS-1 (60%), sem correlação com a expressão proteica, no tempo pós-suplementação com ALA. Portanto, foi demonstrado que o ALA pode modular o ERE através da via da IRE1/XBP, levando ao aumento das chaperonas (BIP/GRP78), além de um efeito adicional na expressão gênica da adiponectina, IL-6 e IRS-1, o que pode demonstrar um potencial terapêutico do ALA em pacientes com DM2. / Type 2 diabetes mellitus (T2DM) is a state of chronic inflammation and activation of endoplasmic reticulum stress (ERS). In this context, studies are necessaries to find new possibilities to improve this inflammation such as the n-3 polyunsaturated fatty acid (n-3 PUFA) acting as an anti-inflammatory agent. In this study, we aimed to evaluate the effect of n-3 PUFA alpha-linolenic acid (ALA, a n-3 PUFA) supplementation in T2DM patients on the molecular expression of ERS genes in abdominal subcutaneous adipose tissue (SAT). We performed a placebo-controlled study, in a double-blind design with 20 T2DM patients, receiving, randomly, 3g/day of ALA or placebo for 60 days. The adipose tissue was collected by fine-needle aspiration in abdomen before and after the supplementation and the genes and proteins were evaluated by real-time PCR and western blot. It was seen, after the supplementation, a reduction in XBP1 (20%), sXBP1 (70%) and an increase in Grp78 (150%) gene expression, likewise same results in protein concentration. Furthermore, it was observed an increase in adiponectina (90%) gene expression and reduction in IL-6 (80%) and IRS-1 (60%) gene expression, with no correlation to protein expression after supplementation of ALA. Therefore, we have provided evidence that ALA may modulate ERS by the IRE1/XBP pathway leading to an increase in chaperones (BIP/GRP78), additionally its effect on adiponectina, IL-6 and IRS-1 gene expression can demonstrate a therapeutic potential in T2DM.

Vu le caractère non-invasif des procédures d’électrophysiologie interventionnelle, la visualisation de régions anatomiques d'intérêt et une orientation adéquate en cours de procédure sont nécessaires. L'objectif de cette thèse est d'étudier, d'optimiser et d'étendre l’utilisation des modalités d'imagerie radiographique. La dose de radiation effective (ED) est calculée d’une façon spécifique au patient pour l’angiographie rotationnelle 3D (3DRA) et il est démontré qu'en ajustant l’acquisition en 3DRA et en fluoroscopie, une réduction de dose importante est possible sans compromettre la qualité d’image nécessaire à la procédure. Un protocole d'acquisition et post traitement pour obtenir une imagerie dynamique basée sur le 3DRA est présenté,permettant une réduction du bruit d'image et une segmentation d'images automatique. L'extraction d'informations dynamiques, structurelles et fonctionnelles à partir d'images MDCT, relatives à la gestion de la fibrillation auriculaire (FA) est étudiée. La fonction auriculaire globale est examinée et des cartes de mouvement régional et de tissu adipeux épicardique sont produites et liés à la FA à différents stades. Une méthode automatisée est présentée pour mesurer l’orifice de l’appendice auriculaire gauche au long du cycle cardiaque et pour optimiser le déploiement de dispositifs de fermeture. Optimiser l’usage des rayons X, les protocoles d'acquisition et les méthodes de post traitement d’images, permet d’obtenir des informations supplémentaires pertinentes aux procédures d'électrophysiologie interventionnelle depuis les modalités d'imagerie radiographiques sans compromettre la qualité d’image ou le flux de travail procédural. / Due to their non-invasive character, interventional electro physiological procedures require visualisation of anatomical regions of interest and adequate guidance of procedural manoeuvres. The aim of this thesis was to study, optimise and expand the use of radiographic imaging modalities. The first part focuses on the influence of C-arm system image acquisition parameters on radiation dose incurred by the patient. We developed a patient-specific way to calculate effective dose (ED) in 3Drotational angiography (3DRA) and showed in 3DRA and fluoroscopy that by applying adequate protocol adjustments, an important dose reduction could be obtained without compromising necessary image quality. The second part focuses on the development and validation of an acquisition and post-processing protocol for dynamic imaging using 3DRA. This method enables automatic image noise reduction and image segmentation. The third part focuses on the extraction of dynamic,structural and functional information from MDCT images, relevant to management of atrial fibrillation (AF). We studied atrial function and generated maps of regional atrial motion and epicardial adipose tissue and related them to AF burden. We also developed an automated method to measure LA appendage orifice dimensions through out the cardiac cycle to optimise measurements for deployment of closure devices.Overall we demonstrate that by optimising radiation usage, acquisition protocols and image post-processing methods, additional information relevant to interventional electrophysiological procedures can be extracted from radiographic imaging modalities without compromising image quality or procedural workflow.

Multipotent mesenchymal stromal cells (MSC) were first discovered in bone marrow and can be isolated from “virtually all organs”. They could participate in tissue maintenance and self-renewing process. They are able to adhere to plastic surfaces and acquire a fibroblastic shape when isolated. They are characterized by a particular phenotype and are able to differentiate into several cell types if cultivated in a specific induction medium. These characteristics were defined on MSC in culture and do not represent how they may be in situ.MSC present particular properties. They can secrete growth factors and several cytokines that give them a trophic activity on one hand and the ability to modulate the immune system on the other hand. They are also able to differentiate. These different properties make them an attractive candidate for cell therapy.MSC are already the focus of several pre-clinical and clinical studies. Nevertheless, the results of these studies are difficult to interpret due to limited understanding of their basic biology. MSC are poorly defined in situ and are heterogeneous. Their heterogeneity is dictated by their tissue of origin and cell preparation. To date, there is no standard protocol for MSC isolation and culture. This leads to numerous questions regarding patient safety, and these questions require answers.The first part of the work deals with the methods used to optimize the extraction of MSC and purification from adipose tissue, one of the main sources of autologous MSC with bone marrow. Classical methods require an enzymatic digestion step. The enzyme used and the duration of adipose tissue digestion time can induce cellular alterations and modify cell functions. Moreover, the addition of a xenobiotic increases the risk of contamination and complicates the monitoring of good manufacturing practices (GMP). We propose a method that does not require this enzymatic digestion step while being easier, safer, faster, gentler and less expensive. Compared to the classical enzymatic method, our method yields an equivalent number of MSC from adipose tissue while preserving their properties.The second part of this work focuses on the characterization of the MSC subpopulations from adipose tissue and compares them to those from bone marrow, which are the historical gold standard. The study made it possible to deepen the knowledge of MSC surface markers in situ from these 2 sources. It also evaluated the various properties of the isolated subpopulations thanks to the cell surface markers CD271, SUSD2, MSCA-1, CD44 and CD34. We showed that MSC from bone marrow express MSCA-1, CD271 and SUSD2 markers in situ. We also found that a population clearly positive for the CD34 does exist in situ with different properties compared to those of the unselected populations or the negative counterpart. 2 populations that are negative and positive for CD44 also exist with similar properties.In contrast to bone marrow MSC, only one selection was able to effectively isolate MSC from adipose tissue by a positive selection based on the expression of CD34. We also isolated a CD271+ population but only from lipoaspirate samples and not from abdominoplasty samples. Collectively, our results suggest that MSCA-1 seems to be the best marker through which to isolate MSC from bone marrow and that CD34 is the only marker able to positively isolate cells from adipose tissue. Thus, we show that the MSC from the different sources share similar properties although they have specific characteristics. The choice of the source and of the marker with which to isolate a particular subpopulation is important depending on their intended clinical use. / Les cellules stromales mésenchymateuses multipotentes (CSM) ont été mises en évidence dans la moelle osseuse et peuvent être isolées de « virtuellement tous les organes ». Elles participeraient à la maintenance et au renouvellement des tissus. Une fois isolées, elles sont capables d’adhérer à des surfaces en plastique en prenant une forme fibroblastique. Elles sont caractérisées par un phénotype particulier et peuvent se différencier en divers types cellulaires lorsque cultivées dans un milieu d’induction spécifique. Ces caractéristiques ont été définies sur les CSM en culture et ne reflètent pas forcément ce qui se passe in situ.Les CSM présentent des propriétés particulières. Elles peuvent sécréter des facteurs de croissance ainsi que de nombreuses cytokines qui leur permettent d’une part d’avoir une activité trophique et d’autre part de moduler le système immunitaire. Elles sont aussi capables de se différencier. Ces différentes propriétés les rendent particulièrement attractives pour la thérapie cellulaire.Les CSM font déjà l’objet de nombreuses études pré-cliniques et cliniques dont les résultats sont difficilement interprétables car nous n’avons à l’heure actuelle qu’une compréhension limitée de leur biologie de base. Les CSM sont encore mal définies in situ et sont hétérogènes. Cette hétérogénéité provient de leur différence d’origine et de leur préparation cellulaire :il n’existe aucune standardisation des protocoles d’isolation et de culture. Cette hétérogénéité entraine de nombreuses questions relatives à la sécurité du patient qui doivent être élucidées.La première partie de ce travail cherche à optimiser les méthodes d’extraction et de purification des CSM du tissu adipeux humain, la principale source de CSM autologues avec la moelle osseuse. Les méthodes classiques requièrent une étape de digestion enzymatique dont l’enzyme utilisée et le temps de digestion du tissu adipeux peuvent induire des altérations cellulaires et modifier leurs fonctions. De plus, l’adjonction de xénobiotiques augmente le risque de contamination et complique le suivi des bonnes pratiques de fabrication (BPF). Nous proposons une méthode qui s’affranchit de cette étape de digestion enzymatique tout en étant plus facile, plus sûre, plus rapide, moins chère et moins traumatisante pour les cellules. Elle permet d’obtenir un nombre tout aussi important de CSM du tissu adipeux que la méthode enzymatique classique en préservant leurs propriétés.La deuxième partie de ce travail vise à caractériser les sous populations de CSM du tissu adipeux humain en les comparant à celles de la moelle osseuse, source de référence historique. Cette étude a permis d’approfondir la connaissance des marqueurs de surface des CSM de ces 2 sources in situ, tout en évaluant les différentes propriétés des sous-populations isolées grâce aux marqueurs de surface CD271, SUSD2, MSCA-1, CD44 et CD34. Nous avons montré que les CSM de la moelle osseuse expriment les marqueurs MSCA-1, CD271 et SUSD2 in situ et qu’il existait une sous-population clairement positive pour le CD34 avec des propriétés différentes de celles de la population non sélectionnée ou négative pour ce marqueur. Il existe aussi 2 sous-populations positive et négative pour le CD44 avec des propriétés similaires.Contrairement aux CSM de la moelle osseuse, une seule sélection a permis d’isoler efficacement les CSM du tissu adipeux par une sélection positive sur base de l’expression du CD34. Nous avons pu aussi isoler une population CD271+ mais seulement des prélèvements de lipoaspirations et non des abdominoplasties.Au vu de nos résultats, MSCA-1 semble le meilleur marqueur pour isoler les CSM de la moelle osseuse tandis que le CD34 est le seul marqueur capable d’isoler positivement celles du tissu adipeux. Ainsi, nous montrons que les CSM issues de différentes sources partagent des propriétés similaires avec cependant des caractéristiques propres. Le choix de la source et du marqueur pour isoler une sous-population sont donc importants en fonction de leur utilité clinique envisagée. / Doctorat en Sciences biomédicales et pharmaceutiques (Médecine) / info:eu-repo/semantics/nonPublished

Abstract Collagen XVIII is an evolutionarily conserved, ubiquitously-expressed basement membrane (BM) proteoglycan produced in three isoforms, the individual roles of which are largely unknown. The physiological in vivo roles of these collagen XVIII isoforms are studied here using novel genetically modified mouse strains deficient in either the short or the medium/long isoforms of the molecule. In addition, the effects of keratin-14-driven overexpression of the thrombospondin-1 (Tsp-1) –like domain, which is common to all three collagen XVIII isoforms, are studied. The findings underline the importance of the short collagen XVIII isoform in the eye, as its absence was sufficient to cause the aberrant vascularisation of the retina previously reported in mice lacking all isoforms of collagen XVIII. In addition, an excess of the collagen XVIII Tsp-1 domain led to serious eye abnormalities, possibly by interfering with the functions of the full-length collagen XVIII produced in mice. Collagen XVIII was also shown to contribute to adipogenesis in an isoform-specific manner, in that a lack of the medium/long isoforms of collagen XVIII led to impaired adipocyte maturation and the subsequent reduction in the adipocyte number induced liver steatosis and hypertriglyceridaemia. Hence this work establishes a new extracellular matrix ECM-directed mechanism contributing to control over the multistep adipogenesis programme and points to the functional consequences of its impairment for ectopic fat deposition. The enzymatic remodelling of ECM components results in molecules with novel biological activities. Arresten is a collagen IV(α1)-derived fragment with anti-angiogenic properties which was originally described as not having any direct anti-tumour effects on cancer cells themselves. The present data revealed novel inhibitory roles for arresten in oral squamous carcinoma cell proliferation, survival, motility and invasion. Since arresten is a potent inhibitor of angiogenesis, the data generated here further underline the possibility for using it as a therapeutic agent in cases of cancer. / Tiivistelmä Kollageeni XVIII on tyvikalvojen proteoglykaani ja yksi harvoista evoluutiossa konservoituneista kollageeneista. Se esiintyy elimistössä kolmena isomuotona, joiden biologiset tehtävät ovat vielä jokseenkin epäselviä. Tässä tutkimuksessa selvitettiin kollageeni XVIII:n isomuotojen fysiologista merkitystä hyödyntäen uusia hiirilinjoja, joilta kollageeni XVIII:n lyhyt tai kaksi pisintä varianttia oli geneettisesti inaktivoitu. Poistogeenisten hiirimallien rinnalle tehtiin kaikille varianteille yhteistä trombospondiini-1 (Tsp-1)-domeinia yli-ilmentävä hiirilinja. Tämän väitöskirjatutkimuksen avulla saatiin uutta tietoa kollageeni XVIII:n ja etenkin sen lyhimmän variantin tärkeästä roolista silmässä. Aikaisemmat tutkimukset ovat osoittaneet kollageeni XVIII:n puutteen häiritsevän silmän verkkokalvon verisuonituksen normaalia kehittymistä. Tässä työssä havaittiin, että pelkästään lyhyen isomuodon puute riitti altistamaan hiiret muutoksille verkkokalvon suonituksessa. Tsp-1-osan ylimäärän havaittiin lisäksi alistavan hiiret muutoksille silmän rakenteessa, mahdollisesti häiritsemällä silmässä jo olemassa olevan kollageeni XVIII:n toimintaa. Tässä työssä havaittiin myös uusi yhteys kollageeni XVIII:n ja rasvasolujen kypsymisen välillä. Verrokkihiiriin verrattuna muodostuvan rasvakudoksen havaittiin jäävän merkittävästi vähäisemmäksi poistogeenisillä hiirillä, joilta kollageeni XVIII:n pitkät isomuodot olivat geneettisesti inaktivoitu. Heikentynyt rasvakudoksen muodostuminen lisäsi triglyseridien kertymistä hiiren verenkiertoon ja maksaan. Tutkimustulos on merkittävä avaus soluväliaineen merkityksestä rasva-aineenvaihdunnalle ja kannustaa lisätutkimuksilla selvittämään, onko kollageeni XVIII:lla yhteys myös ihmisen metaboliseen oireyhtymään. Soluväliaineen komponenttien entsymaattinen muokkaus tuottaa usein molekyylejä, joilla on uusia isäntämolekyyleistä poikkeavia ominaisuuksia. Tässä työssä tutkittiin yhden tällaisen molekyylin, tyvikalvokollageenin IV hajoamistuotteen, arrestenin, suoria vaikutuksia syöpäsoluille. Arrestenin tiedettiin entuudestaan estävän syöpäkasvainten verisuonten uudismuodostusta koe-eläimillä. Työssä osoitettiin, että arresten vaikutti endoteelisolujen lisäksi myös itse syöpäsoluihin estäen niiden lisääntymistä ja vähentäen niiden elinkykyä ja liikkuvuutta, mikä tekee arrestenista entistä houkuttelevamman ehdokasmolekyylin lääkekehitystyöhön.