Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-07-19T14:58:19Z No. of bitstreams: 1 dianemichelaneryhenrique.pdf: 7655862 bytes, checksum: 28c817a6bfd0f9b2a6f1809cbed1cc96 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-08-09T12:19:37Z (GMT) No. of bitstreams: 1 dianemichelaneryhenrique.pdf: 7655862 bytes, checksum: 28c817a6bfd0f9b2a6f1809cbed1cc96 (MD5) / Made available in DSpace on 2017-08-09T12:19:37Z (GMT). No. of bitstreams: 1 dianemichelaneryhenrique.pdf: 7655862 bytes, checksum: 28c817a6bfd0f9b2a6f1809cbed1cc96 (MD5) Previous issue date: 2009-03-13 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Na população geral, a prática regular de exercícios físicos se associa à melhora da capacidade funcional e à redução de eventos cardiovasculares. Por outro lado, em portadores de doença renal crônica, uma população que apresenta significativo comprometimento da capacidade funcional, alta prevalência de hipertensão arterial e elevadas taxas de mortalidade cardiovascular, poucos são os estudos que avaliam o efeito da atividade física na capacidade funcional e no controle da hipertensão arterial. Portanto, o objetivo deste estudo foi avaliar o efeito do treinamento aeróbico, realizado durante as sessões de hemodiálise, sobre a capacidade funcional e sobre a pressão arterial de pacientes renais crônicos. Foram avaliados 14 indivíduos portadores de doença renal crônica sob tratamento hemodialítico, antes e depois de 12 semanas de treinamento aeróbico realizado em cicloergômetro de membros inferiores, durante as sessões de hemodiálise. Constatou-se que dentre os pacientes, quatro eram homens e 10 eram mulheres, com média de idade de 47,6 ± 12,79 anos, cinco eram brancos e nove eram negros e que o tempo de hemodiálise variou de 93,7 ± 43,90 meses. Os pacientes foram submetidos à monitorização ambulatorial da pressão arterial de 24 horas, ao teste de caminhada de 6 minutos e ao teste cardiopulmonar de exercício tanto antes como depois do período de treinamento (p < 0,05). Após o período de treinamento aeróbico, observou-se aumento da distância percorrida no teste de caminhada de 6 minutos de 509 ± 91,9 m para 555 ± 105,8 m (p < 0,001), além de redução da pressão arterial sistólica de 151 ± 18,4 mm Hg para 143 ± 14,7 mm Hg (p = 0,08), pressão arterial diastólica de 94 ± 10,5 mm Hg para 91 ± 9,6 mm Hg (p = 0,05), pressão arterial média de 114 ± 13,0 mm Hg para 109 ± 11,4 mm Hg (p = 0,07), pressão arterial sistólica no sono de 150 ± 23,9 mm Hg para 140 ± 19,3 mm Hg (p = 0,02), pressão arterial diastólica no sono de 93 ± 15,3 mm Hg para 88 ± 14,3 mm Hg (p = 0,01), pressão arterial média no sono de 112 ± 18,0 mm Hg para 106 ± 16,1 mm Hg (p = 0,04). O valor de pico de consumo de oxigênio no primeiro e segundo limiar ventilatório aumentou de 20,6 ± 6,92 mUkg.m-1 para 21,2 ± 10,13 milkg.m-1 (p = 0,45) no pré e pós-treinamento aeróbico. Nas variáveis laboratoriais a hemoglobina aumentou de 10,8 ± 1,20 g/dL para 11,6 ± 11,6 g/dL (p = 0,04), o hematócrito aumentou de 32,8 ± 3,57% para 35,2 ± 2,76 (p = 0,03) e os triglicérides aumentou de 96,6 ± 35,17 mg/dL para 127,0 ± 54,56 (p = 0,04) no pré e pós-treinamento aeróbico. Com base nos resultados obtidos pode-se concluir que o treinamento aeróbico realizado durante as sessões de hemodiálise contribuiu para a melhora da capacidade funcional e para o melhor controle da hipertensão arterial de pacientes portadores de doença renal crônica. / In general population, physical fitness has been shown to improve functional capacity and to reduce cardiovascular events. On the other hand, in patients with chronic kidney disease, which have significant reduction of functional capacity, high prevalence of hypertension and high rates of cardiovascular mortality, there are only a few studies evaluating the effects of exercise training on functional capacity and on blood pressure control. Therefore, the objective this study was to evaluate the effect of aerobic training performed during hemodialysis sessions on functional capacity and hypertension control of patients with chronic kidney disease. Fourteen patients with chronic kidney disease on hemodialysis treatment were evaluated before and after 12 weeks of aerobic training cycle held in the lower limbs during the hemodialysis sessions. Patients underwent 24 hours ambulatory blood pressure monitoring, six-minute walking test and exercise cardiopulmonary test before and after the period of 12 weeks of training (p < 0,05). It was found that among the patients, four males and 10 females, mean age 47,6 ± 12,79 years, five were white and nine were black and the time of hemodialysis ranged from 93,7 ± 43,90 months. The patients underwent ambulatory blood pressure monitoring, 24 hours, the test for 6 minutes of walking and exercise cardiopulmonary test both before and after the period of training (p < 0.05). After the period of aerobic training, there is increasing distance in the test of the 6 minute walk 509 m to 555 ± 91.9 ± 105.8 m (p < 0, 001), and reduction of systolic blood pressure of 151 ± 18.4 mm Hg to 143 ± 14,7 mm Hg (p = 0,08), diastolic blood pressure of 94 ± 10,5 mm Hg to 91 ± 9,6 mm Hg (p = 0,05), pressure mean blood of 114 ± 13.0 mm Hg to 109 ± 11.4 mm Hg (p = 0,07), systolic blood pressure in the sleep of 150 ± 23,9 mm Hg to 140 ± 19,3 mm Hg (p = 0,02), diastolic blood pressure in the sleep of 93 ± 15,3 mm Ng to 88 ± 14,3 mm Ng (p = 0,01), mean arterial pressure during sleep was 112 ± 18.0 mm Hg to 106 ± 16.1 mm Hg (p = 0,04). The value of peak oxygen consumption in the first and second ventilatory threshold increased from 20,6 ± 6,92 mL/kg.m-1 to 21,2 ± 10,13 mL/kg.m-1 (p = 0,45) in pre-and post-aerobic training. Laboratory variables in the hemoglobin increased from 10,8 ± 1.20 g/dL to 11,6 ± 11,6 g/dL (p = 0,04), the hematocrit increased from 32,8 ± 3,57% to 35,2 ± 2,76 (p = 0,03) and triglycerides increased from 96,6 ± 35,17 mg/dL to 127,0 ± 54,56 (p = 0,04) in pre-and post-aerobic training. Based on the results we can conclude that aerobic training performed during hemodialysis sessions improved functional capacity and contributed to hypertension control of patients with chronic kidney disease.

Abstract Coronary artery disease is the leading cause of death in the world. The outcome of patients at a very high operative risk undergoing coronary artery bypass surgery has not been thoroughly investigated. Cohorts of patients underwent coronary surgery between January 1997 and December 2013 at the Oulu University Hospital, Finland. Data was acquired from electronic patient records. Statistical analysis was performed on the collected data to evaluate outcome and identify predictors of adverse events. Very high-risk patients who underwent isolated coronary artery bypass surgery had a high 30-day mortality (16.2%), but their 5-year survival was satisfactory (66.8%). Survivors of out-of-hospital cardiac arrest were compared to a control group. Immediate postoperative mortality was slightly higher in out-of-hospital cardiac arrest patients (6.3% vs. 0%, p = 0.24), but the overall 5-year survival rates were similar (80.7% vs. 84.5%). Patients with preoperative stage 3 chronic kidney disease have a higher mortality than patients with stage 1-2 chronic kidney disease. Kidney function decline/year was predictive of all-cause mortality, cardiovascular mortality and also tended to predict fatal and non-fatal cardiovascular events. The E-CABG postoperative complication grading system was used to stratify the severity and prognostic impact of postoperative complications and was shown to predict early and late mortality for these patients. The outcome of emergency coronary artery bypass surgery was studied in a multi-center setting. Increasing emergency classes, left ventricular ejection fraction ≤30%, on-pump surgery, and participating centers were independent predictors of in-hospital mortality. Survival rates at 1, 3 and 5 years were 86.4%, 81.6%, and 76.1%. Despite the high preoperative risk of these patients, the long-term outcome for coronary surgery is satisfactory. Patients with stage 3 chronic kidney disease may experience a significant decline in kidney function and poor outcome. Early referral to a nephrologist may be beneficial for these patients. The E-CABG complication grading system seems to be a promising tool for stratifying the severity and prognostic impact of complications occurring after coronary surgery. / Tiivistelmä Sepelvaltimotauti on johtavia kuolinsyitä Maailmassa. Ohitusleikkauksen tuloksia ei ole täysin selvitetty erittäin korkean riskin potilailla. Potilaat leikattiin vuosina 1997-2013. Potilastiedot hankittiin sairauskertomuksista ja kuolinsyytiedot kansallisista rekistereistä. Erittäin korkean riskin potilaiden välitön kuolleisuus ohitusleikkauksen jälkeen on korkea (30 päivän kuolleisuus 16,2 %). Viiden vuoden kuluttua leikkauksesta elossa oli 66,8% leikatuista. Ohitusleikkausta edeltävästi elvytettyjä potilaita verrattiin kontrolliryhmään. Välittömät leikkauksen jälkeinen kuolleisuus oli 6,3% vs. 0% (p = 0,24). Viiden vuoden kuluttua leikkauksesta elossa oli tutkimusryhmästä 80,7% ja kontrolliryhmästä 80,7%. Leikkausta edeltävästi keskivaikean munuaisten vajaatoiminnan omaavilla potilailla on korkeampi kuolleisuus verrattuna potilaisiin, joiden munuaistoiminta on normaalia tai lievästi heikentynyt. Munuaisten vajaatoiminnan eteneminen ennusti kokonaiskuolleisuutta, sydän- ja verisuonikuolleisuutta ja enteili sydän- ja verisuonitapahtumia. E-CABG leikkauksen jälkeisten komplikaatioiden luokittelujärjestelmällä luokiteltiin leikkauksen jälkeisten komplikaatioiden vaikeusastetta ja ennusteellista vaikutusta. E-CABG luokat ja pisteytys ennustivat 1kk, 3kk kuolleisuutta ja kuolleisuutta pidemmällä aikavälillä. Päivystysohitusleikkauksen tuloksia tutkittiin monikeskusasetelmassa. Sairaalakuolleisuutta ennustivat päivystysleikkausluokitteluluokan vakavuus, vasemman kammion ejektiofraktio ≤30%, perfuusiossa tehty leikkaus ja leikkaava keskus. Potilaiden elossaololuvut olivat 1, 3 ja 5 vuoden kohdalla 86,4%, 81,6%, and 76,1%. Leikkaustulokset erittäin korkean riskin potilailla ohitusleikkauksesta ovat kohtuullisia leikkausta edeltävään riskiarvioon suhteutettuna. Näin ollen tämän potilasryhmän sepelvaltimotaudin hoito leikkaamalla on perusteltua. Keskivaikean munuaisten vajaatoiminnan omaavien potilaiden munuaissairauden etenemiseen seuranta-aikana liittyy kuolleisuutta ja sydän- ja verisuonitapahtumia. Aikaisessa vaiheessa tehty nefrolgin konsultaatio voi parantaa näiden potilaiden munuaisfunktiota. E-CABG komplikaatioiden luokittelujärjestelmä vaikuttaa lupaavalta työkalulta ohitusleikkauksen jälkeisten komplikaatioiden luokitteluun ja ennustevaikutuksien arviointiin.

La polykystose rénale autosomique dominante (ADPKD) est la maladie monogéniquehumaine la plus fréquente (prévalence 1/800). Les gènes responsables de cette maladie sont PKD1(codant pour PC1) ou PKD2 (codant pour PC2). La maladie évolue vers l’insuffisance rénale terminale.Aujourd’hui, seul un traitement symptomatique est proposé aux malades. Les mécanismes à l’originede l’ADPKD sont mal connus. Les modèles animaux permettent de mieux comprendre laphysiopathologie d’une maladie. Il n’existe pas de bon modèle de polykystose (même causemoléculaire, même mode de transmission, même signes cliniques). En utilisant la transgénose degrands fragments, nous avons créé un modèle de surexpression de PKD2 humain. Le transgène estsous le contrôle de son promoteur naturel humain. Cette souris exprime deux fois plus de PC2 queles sauvages. Elle ne présente que quelques microkystes mais une tubulopathie associant défaut deconcentration des urines et protéinurie tubulaire. La surexpression de PC2 inhibe l’expression degènes codant pour des protéines de la matrice extracellulaire. Le phénotype cellulaire de cesanimaux est remarquable : un tiers des cellules présentent un nombre élevé de centrosomes. Cephénotype cellulaire a été retrouvé chez des souris sous exprimant Pkd2 et chez des souris sousexprimant Pkd1. Ce caractère multicentrosomique est corrigé en incubant les cellules avec uninhibiteur de CaMKII ou en croisant nos souris transgéniques avec une souris KO de Camk2. Nousavons réussi à lier CaMKII, la duplication des centrosomes et les polycystines, in vitro et in vivo. Ceciamène un éclairage nouveau sur la duplication du centrosome et la physiopathologie de l’ADPKD. / Autosomal Dominant Polycystic Kidney Disease (ADPKD) is the most common monogenic human disease (prevalence 1/800). Genes responsible for this disease are PKD1 (encoding PC1) or PKD2 (encoding PC2). The disease progresses to end stage renal disease. Today, only symptomatic treatment is offered to patients. The mechanisms underlying the ADPKD are unknown. Animals models allow better understand the disease’s pathophysiology. There is no good model of ADPKD (same molecular cause, same clinical signs). We created a mice model of human PKD2 overexpression. The transgène is under the control of its human natural promoter. This mouse expresses PC2 twice as much as the wild. It shows only few microcysts but tubulopathy involving lack of urine concentration and tubular proteinuria. PC2 overexpression inhibits the expression of genes encoding proteins of the extracellular matrix. The cellular phenotype of these animals is special : one third of the cells have a high number of centrosomes. This cellular phenotype was found in Pkd2 Knockout mice and in Pkd1 knockout mice. This multicentrosomic character is corrected by incubating the cells with a CaMKII inhibitor or by crossing our transgenic mice with Camk2 knockout mice. We propose a link between CaMKII, Centrosome duplication and polycystin in vitro and in vivo. This brings a new light on centrosome duplication and pathophysiology of ADPKD.

Le groupe des KDIGO a défini un nouveau syndrome regroupant les troubles minéraux et osseux et les calcifications vasculaires au cours de la maladie rénale chronique {TMO-MRC). Les TMO-MRC sont associés à une augmentation du risque de fracture, de calcification vasculaire et à une surmortalité. La quête d'un biomarqueur fiable de la maladie osseuse et des calcifications vasculaires reste le défi du néphrologue. Au cours des dernières années, de nombreuses protéines de l'os ont été associées aux calcifications vasculaires chez les patients urémiques, tels que les phosphatases alcalines osseuses et, plus récemment, la sclérostine. Cette petite protéine est secrétée par l'ostéocyte et inhibe l'ostéoformation en bloquant la voie de signalisation Wnt dans l'ostéoblaste. Il a récemment été suggéré que la sclérostine aurait une activité catabolique sur l'os et serait impliquée dans la déminéralisation du squelette. L'objectif de ce travail était d'étudier la sclérostine au cours de la MRC. Nous avons tout d'abord montré que les concentrations sériques de sclérostine augmentaient avec la baisse du débit de filtration glomérulaire mesurée par la clairance de l'inuline et ce dès le stade 3 de la MRC, indépendamment de l'âge. De plus, cette étude nous a permis de montrer pour la première fois que la phosphorémie était indépendamment et positivement associée à la sclérostine sérique. Ensuite, nous avons retrouvé une association positive et forte entre la concentration sérique de sclérostine et les calcifications vasculaires chez des patients en hémodialyse chronique. Enfin, nous avons montré que les calcifications artérielles étaient significativement associées à une qualité osseuse corticale altérée étudiée en scanner quantitatif de haute résolution. Ces résultats suggèrent que la sclérostine pourrait constituer un messager important dans la relation entre l'os et la paroi vasculaire calcifiée des patients atteints d'une insuffisance rénale chronique terminale / The KDlGO group identified a new syndrome involving mineral and bone disorders and vascular calcification in chronic kidney disease (CKD-MBD}. CKD-MBD are associated with an increased risk of fracture, vascular calcification and increased mortality. The search for a reliable biomarker of bone disease and vascular calcification remains the challenge of the nephrologist. ln recent years, many bone proteins have been associated with vascular calcification in uremic patients, such as bone alkaline phosphatase and more recently sclerostin. This small protein is secreted by the osteocyte and is an inhibitor of bone formation by blocking the Wnt signaling in osteoblasts. lt has recently been suggested that sclerostin has a catabolic activity on bone and is involved in the demineralization of the skeleton. The aim of this research was to study the sclerostin during the course of CKD. We first showed that serum levels of sclerostin increased with the decrease of glomerular filtration rate measured by inulin clearance already from stage 3 chronic kidney disease, regardless of age. Furthermore, in this study, we showed for the first time that serum phosphorus was independently and positively associated with serum sclerostin. Subsequently we found a positive and strong association between serum sclerostin and vascular calcification in maintenance hemodialysis patients. Finally, we have shown that arterial calcification were significantly associated with an altered cortical bone quality studied by high resolution peripheral quantitative computed tomography. These results suggest that sclerostin could be an important messenger in the cross-talk between the bone and the calcified vascular wall in end stage renal disease

Dans un contexte de vieillissement de la population et d’accroissement des maladies chroniques liées à l’âge comme le diabète, de nouveaux biomarqueurs de l’état de santé à long terme doivent être étudiés. Les produits de glycation avancée (AGE) sont des molécules témoins de la charge métabolique accumulée au cours du temps, dénommée "mémoire métabolique". Les AGE jouent un rôle important dans les lésions à long terme dans le diabète et dans le déclin du métabolisme global lié au vieillissement. L’accumulation cutanée des AGE peut être mesurée par autofluorescence (AF) de manière instantanée et non invasive grâce à l’AGE-READER. Les objectifs de cette thèse étaient d’évaluer la valeur de l’AF cutanée des AGE en tant que marqueur de mémoire métabolique chez des personnes âgées de la cohorte des 3-Cités et parallèlement d’évaluer la valeur pronostique de l’AF pour les complications du diabète chez des patients porteurs de diabète de type 1. Chez les personnes âgées, nous avons montré que l’AF reflétait les statuts glycémique et rénal 10 ans avant la mesure. Chez les patients atteints de diabète de type 1, l’AF était associée à la présence d’une neuropathie 4 ans plus tard. De plus, dans cette même population, nous avons décrit l’évolution de l’AF sur 4 ans de suivi. Nous avons montré que les principaux déterminants de son évolution étaient la fonction rénale et le traitement par pompe à insuline. Enfin nous avons trouvé que l’augmentation de l’AF sur 4 ans de suivi était associée à la survenue de la maladie rénale. Ces travaux soulèvent de nouvelles perspectives de recherche quant à l’intérêt de l’AF à différents âges clés de la vie en tant que biomarqueur de pathologies qui évoluent sur des dizaines d’années. / In the context of the ageing of the population and the increase of age related diseases such as diabetes, new biomarquers of the long-term health status should be considered. Advanced glycation end products (AGE) are molecules indicators of the metabolic burden over time, called “metabolic memory”. AGE play an important role in long term diabetes injuries and in the global decline of the metabolism related to ageing. Skin accumulation of AGE can be measured by autofluorescence instantly and non-invasivly with a tool called AGE-READER. The objectives of my dissertation were to evaluate the value of the skin autofluorescence (sAF) of AGE as marker of metabolic memory in elderly people from the 3-City cohort and in parallel, in patients with type 1 diabetes, evaluate the prognostic value of sAF for diabetes complications. In the elderly population, we showed that sAF reflected glycemic and renal status of 10 years before. In patients with type 1 diabetes, sAF was associated to the presence of neuropathy 4 years later. Moreover, in this same population, we described the evolution of sAF in 4 years of follow-up and we showed that the principal determinants of the evolution of sAF were kidney function and insulin pump therapy. Finally, we also found that increase of sAF in 4 years was associated with the occurrence of kidney disease. This work rises new research opportunities about the interest of sAF at differents key ages as biomarker of pathologies which evolve in several decades.

Submitted by Nadir Basilio (nadirsb@uninove.br) on 2018-06-19T17:48:13Z No. of bitstreams: 1 Nina Teixeira Fonseca.pdf: 4441067 bytes, checksum: c9f9edd5e35ac829504bbbe923461f01 (MD5) / Made available in DSpace on 2018-06-19T17:48:13Z (GMT). No. of bitstreams: 1 Nina Teixeira Fonseca.pdf: 4441067 bytes, checksum: c9f9edd5e35ac829504bbbe923461f01 (MD5) Previous issue date: 2015-12-16 / Introduction: Currently, chronic kidney disease (CKD) is one of the most serious public health problems, becoming a global epidemic. It is also known that the volume of the rostral fluid displaced during the night, from the lower limbs is associated with increased neck circumference and severity of obstructive sleep apnea (OSA) in CKD patients undergoing hemodialysis (HD). Objectives: To determine the prevalence of excessive daytime sleepiness, the degree of collapsibility of the upper airways (UA) through the negative expiratory pressure (NEP) test in patients with CKD undergoing HD. Methods: A cross-sectional clinical study involving patients with CKD undergoing HD. It used the Epworth Sleepiness Scale and held the NEP test before and after the performance of the HD sessions. Results: It was observed that 70% of CKD patients undergoing HD exhibit excessive daytime sleepiness. The mean circumference of the neck pre dialysis was 38.60 ± 4.32 cm whereas after dialysis was 37.92 ± 3.89 cm and the weight of the pre hemodialysis patients had an average of 72.4 ± 21 15 kg and 70.64 ± 21.21 kg after hemodialysis. The average value V0,2 was 19.98 ± 11.77% before dialysis and 28.60 ± 21.98% after dialysis. Conclusion: Patients with CKD undergoing HD have excessive daytime sleepiness and increased expiratory flow after the NEP test. The NEP test can be stated as intervention assessment tool along the upper airway in the direction of increased expiratory flow. / Introdução: Atualmente, a doença renal crônica (DRC) é um dos mais sérios problemas de saúde pública, tornando-se uma epidemia global. Sabe-se também que o volume de fluido rostral deslocado durante a noite, proveniente dos membros inferiores, está relacionada ao aumento da circunferência do pescoço e a gravidade da apnéia obstrutiva do sono (AOS) em pacientes com DRC submetidos a hemodiálise (HD). Objetivos: Verificar a prevalência de sonolência excessiva diurna e o grau de colapsabilidade das vias aéreas superiores (VAS) através do teste de pressão negativa expiratória (PNE) em pacientes com DRC submetidos a HD. Método: Foi realizado um estudo clinico transversal envolvendo pacientes com DRC submetidos a HD. Foi utilizado a escala de sonolência de Epworth e realizado o teste de PNE antes e após a realização das sessões de HD. Resultados: Foi observado que 70% dos pacientes com DRC submetidos a HD apresentam sonolência diurna excessiva. A média da circunferência do pescoço pré hemodiálise foi de 38,60 ± 4,32 cm, enquanto que pós hemodiálise foi de 37,92 ± 3,89 cm e o peso dos pacientes pré hemodiálise apresentou uma média de 72,4 ± 21,15 kg e de 70,64 ± 21,21 kg pós hemodiálise. O valor médio de V0,2 foi de 19,98 ± 11,77% pré diálise e de 28,60 ± 21,98% pós diálise. Conclusão: Pacientes com DRC submetidos a HD apresentam sonolência excessiva diurna e aumento do fluxo expiratório verificado pelo teste de PNE. O teste de PNE pode ser indicado como instrumento de avaliação de intervenções junto as vias aéreas superiores no sentido de aumento do fluxo expiratório.

Estudos tem demonstrado que a injúria renal aguda (IRA) pode evoluir para doença renal aguda ou crônica, principalmente quando associada à comorbidades como a hipertensão sal-dependente. Além disso, o desenvolvimento da perda de função renal é comumente associado a distúrbios mineral e ósseo. O uso de antioxidantes como a N-acetilcisteína (NAC) tem sido utilizado para prevenir a lesão da IRA, no entanto pouco se sabe sobre o uso deste medicamente em um período prolongado e os seus efeitos no metabolismo mineral e ósseo. Sendo assim os objetivos do presente estudo foram avaliar a repercussão da sobrecarga de sódio na dieta em modelo de IRA com ou sem tratamento com NAC, e observar a influência da IRA, dieta hipersódica e tratamento no metabolismo mineral e ósseo após 10 semanas de protocolo. Para isto foram utilizados ratos machos Wistar alimentados com dieta normossódica (0,5% Na - NS) ou hipersódica (3,2% Na - HS), tratados com ou sem NAC (IR NAC) (600mg/L) e submetidos à cirurgia de isquemia (45 minutos) e reperfusão renal bilateral (IR) ou Sham. Os animais foram acompanhados por 10 semanas após a cirurgia. Foram avaliadas pressão arterial caudal, função renal, metabolismo mineral e histomorfometria óssea. Os animais que sofreram IR e foram alimentados com dieta HS apresentaram maior excreção de sódio, cálcio e fósforo e albuminúria. A albuminúria correlacionou-se com o aumento do PTH induzindo a um maior tecido osteoide e superfície de osteoblastos nestes animais. A NAC diminuiu excreção de sódio, cálcio, fósforo, albuminúria e PTH, mas causou a diminuição do volume ósseo e aumento da separação trabecular. A ingestão do sódio influenciou nos níveis séricos altos de 1-25(OH)2D e baixos de FGF-23. Estes resultados sugerem que a sobrecarga de sal na dieta causou evolução da IRA após 10 semanas. Essa perda de função renal aumentou o PTH, volume osteoide e osteoblastos. Em contrapartida, o tratamento com a NAC preveniu a progressão da disfunção renal, mas levou à perda de massa óssea / Studies have shown that acute kidney injury (AKI) can progress to acute or chronic kidney disease especially when associated with co-morbidities such as salt-dependent hypertension. In addition to impairment of renal function there is association with mineral and bone disorders. The antioxidants such as N-acetylcysteine (NAC) has been used to prevent the AKI; however; the prolonged use and its effects in mineral and bone metabolism is not well studied. The aim of the study was to evaluate dietary sodium overload repercussion on the AKI associated to NAC treatment and to observe its effects on bone metabolism after 10 weeks. Were used Wistar rats were fed with normal sodium (0.5% NaCl - NS) or high sodium (3.2% NaCl - HS) diet and treated or not with NAC (600 mg/L) and submitted to renal ischemia (45 minutes) and reperfusion (IR) or Sham surgery. The animals were followed up for 10 weeks after surgery. We evaluated caudal blood pressure, renal function, mineral metabolism and bone histomorphometry. In our results urinary sodium, calcium, phosphorus excretion and albuminuria were higher in HS IR animals. There was a positive correlation between albuminuria and PTH leading to osteoid tissue and surface of osteoblasts augmentation in these animals. In other hand, NAC decreased sodium, calcium, phosphorus excretion, albuminuria and PTH; however, it was observed lower bone volume and higher trabecular separation. Sodium intake had an isolated effect on 1-25(OH)2D and FGF-23. These results suggest that overload salt diet caused AKI development. Loss of renal function increased PTH, osteoid volume and osteoblasts Treatment with NAC prevented renal function impairment, however it induced to bone mass loss

La información sobre las enfermedades renales en mascotas ofrecida en el Hospital de Referencia Dr. Maykel Povea es importante debido a que los usuarios deben conocer el cuidado y prevenir el problema de salud. La presente investigación tiene como objetivo implementar una pieza gráfica que sea fácil de entender para los usuarios y que sirva como complemento en las explicaciones del veterinario dentro del contexto de una cita en el Hospital. Para ello fue usado el cómic, la ilustración y el storytelling como conjunto narrativo y gráfico para presentar la información brindada. Ello fue resuelto de varias maneras. La investigación y entrevistas permitieron reconocer la situación de comunicación entre emisor(hospital) y receptor(usuario). Al mismo tiempo el resultado de la investigación llevó a la creación de varias propuestas gráficas las cuales fueron descartadas según sus valores de ventajas y desventajas. Una vez desarrollada la última propuesta, esta fue sujeta a verificaciones por una serie de prototipos y testeos, en ellos se evaluó: historia, imagen, lectura, recorrido visual, tipografía, etc. Los resultados obtenidos fueron que el storytelling y el cómic son métodos interesantes que intriga y facilita el entendimiento de temas complejos puesto que permiten poner en un contexto similar a los usuarios, lo que les permite sentirse identificados al simpatizar con la historia, con la posibilidad de un aprendizaje significativo. En conclusión, es pertinente continuar la exploración del storytelling como método para presentar temas complejos puesto que es un facilitador para el entendimiento de cualquier tipo de información presentada. / The information on kidney diseases in pets offered at the Dr. Maykel Povea Reference Hospital is important because users must know how to care and prevent this type of health problems. The objective of this research is to implement a graphic piece that is easy for users to understand and to serve as a complement to the veterinarian’s explanations within the context of an hospital appointment. For this, the comic, illustration and storytelling were used as a narrative and graphic set to present the information provided. This was resolved in various ways. The research and interviews made it possible to recognize the communication situation between sender (hospital) and receiver (user). At the same time, the result of the investigation led to create several graphics proposals which were discarded according to their values of advantages and disadvantages. Once the last proposal was developed, it was put as a subject of verification by a series of prototypes and tests, in which the following were evaluated: story, image, reading, visual reading, typography, etc. The results obtained were that storytelling and comics are interesting methods that intrigue and facilitate the understanding of complex issues since they allow users to be put in a similar context, which helps them to feel identified by sympathizing with the story, with the possibility of meaningful learning. In conclusion, it is pertinent to continue the exploration of storytelling as a method to present complex issues since it is a facilitator for the understanding of any type of information presented. / Trabajo de investigación

Hypertension artérielle résistante et maladie rénale chronique : Déterminants et risques associésL’hypertension résistante, définie par une pression artérielle au-dessus de la cible en dépit de la prise de trois antihypertenseurs à dose optimale dont un diurétique, est fréquemment associée à la maladie rénale chronique (MRC). Sa prévalence, ses déterminants et l’impact potentiel de la MRC sur son pronostic à long terme sont mal connus, notamment chez le sujet âgé. Dans l’étude des 3 cités, incluant 4262 personnes de plus de 65 ans traitées pour hypertension, la prévalence de l’hypertension apparemment résistante (HTAR) - la notion de traitement à dose optimale étant inconnue - était de 11,8% vs 5,2% chez ceux avec vs sans MRC (définie par une fonction rénale < 60 mL/min/1.73 m2). Nous avons montré que l’apparition d’une HTAR était plus fortement liée à la rapidité du déclin annuel de la fonction rénale qu’à son niveau, indépendamment des autres facteurs de risque : obésité, diabète, sexe masculin, antécédent cardiovasculaire. Comparé au groupe de référence (avec hypertension contrôlée et sans MRC), les personnes avec une HTAR et une MRC n’avaient pas de risque significativement plus élevé de mortalité toute cause, mais avaient deux fois plus de risque d’accident vasculaire cérébral (AVC), létal ou non, et de récurrence d’un AVC ou d’un événement coronaire, et trois fois plus de décès coronaire. Cependant, l’’hypothèse d’un effet aggravant de la MRC sur le pronostic de l’HTAR n’a pas été confirmé (interaction non significative).Dans la cohorte CKD-REIN, incluant plus de 3000 patients avec une MRC modérée ou avancée suivis en néphrologie (âge moyen, 70 ans, 60% d’hommes), nos résultats préliminaires montrent une prévalence élevée d’HTAR, 36,7%, et plusieurs facteurs de risque potentiellement modifiables : adhérence médiocre au traitement, absence de diurétique, consommation de sel en excès, obésité.Dans l’ensemble, ces travaux montrent l’importance de la MRC dans le développement de l’HTAR et des risques cardiovasculaires associés, et suggère des moyens de prévention au-delà des traitements médicamenteux. / Resistant hypertension and chronic kidney disease: Determinants and outcomesResistant hypertension defined as blood pressure above goal despite simultaneous use of 3 antihypertensive classes at optimal doses including a diuretic, is commonly associated with chronic kidney disease (CKD). Resistant hypertension prevalence and determinants, and the impact of CKD on its long term outcomes are poorly known, particularly in the elderly population.In the 3 Cities cohort, including 4262 community-dwelling elderly individuals, aged 65 years or older treated for hypertension, the prevalence of apparent treatment resistant hypertension (aTRH) – because of lack of information on optimal treatment dose – was 11.8% vs 5.2% in those with vs without CKD (defined as estimated glomerular filtration rate < 60 mL/min/1.73 m2). We showed that new-onset aTRH was more strongly related to the speed of kidney function decline than kidney function level itself, independent of other risk factors: male sex, obesity, diabetes, and history of cardiovascular disease. Compared to the reference group (with controlled hypertension and no CKD), participants with aTRH and CKD had no significantly higher risk of all-cause mortality, but had a risk of fatal or non-fatal stroke and of recurrent stroke or coronary events more than twice as high, and of coronary death more than three times higher. However, the hypothesis that CKD may worsen the prognosis of aTRH was not confirmed (no significant interaction).In the CKDREIN cohort, which included more than 3000 nephrology outpatients with moderate or severe CKD (mean age, 70 years, 60% of men), our preliminary results showed a high prevalence of aTRH, 36,7% and several potentially modifiable risk factors : poor treatment adherence, lack of diuretic use, excess salt intake and obesity.Overall, this work shows the importance of CKD in the development of aTRH and associated cardiovascular outcomes, and suggests means for prevention beyond drug therapy.

Il termine malattia renale cronica (Chronic Kideny Disease: CKD) si riferisce a differenti condizioni caratterizzate da un progressivo declino della funzione renale. Le linee guida internazionali hanno definito la CKD come una condizione in cui siano presenti marcatori di danno renale e/o la velocità di filtrazione glomerulare stimata (Estimated Glomerular Filtration Rtae: eGFR) sia inferiore a 60 ml/min/1.73 m2 per almeno 3 mesi. L’insufficienza renale in stadio terminale è associata ad un alto rischio di malattia cardiovascolare (Cardiovascular Disease: CVD), la più frequente causa di morte in questi pazienti. Fattori di rischio “non-tradizionali” come: infiammazione cronica, stress ossidativo, deplezione proteico-energetica, disordini del metabolismo minerale e deficit di inibitori della calcificazione, partecipano alla patogenesi della CVD. L’infiammazione gioca un ruolo cruciale nella risposta fisiologica all’infezione e al danno renale e partecipa anche nell’evoluzione del danno renale irreversibile con la produzione di diverse molecole infiammatorie a partire da acidi grassi polinsaturi a lunga catena (Long Chain PolyuUsaturated Fatty Acids: LCPUFA) della serie Omega-6. La supplementazione di Omega-3, con effetto antinfiammatorio, nei pazienti affetti da CKD è stata ed è oggetto di molti studi, nonostante ciò, l’effetto sul danno renale è ancora poco chiaro. Comunque, è ampiamente riconosciuto che un alterato profilo lipidico possa determinare la progressione della patologia, inducendo lo stato infiammatorio. Inoltre, elevati/normali livelli di Omega-3 potrebbero essere associati al miglioramento della funzionalità renale, diminuendo quindi il rischio di peggioramento della malattia. Le concentrazioni e il rapporto di Omega-3 e Omega-6 sono strettamente associati alla salute del rene, poiché svolgono ruoli importanti in differenti vie metaboliche. Un altro aspetto, preso poco in considerazione, è l’effetto dei livelli di acidi grassi circolanti e dei loro metaboliti sullo stato infiammatorio e sulla sua modulazione. Il primo scopo di questo studio è stato quello di analizzare il profilo degli acidi grassi in soggetti anziani affetti da CKD. Sono stati arruolati 57 pazienti afferenti agli ambulatori di Nefrologia dell’Ospedale Maggiore Policlinico di Milano e sono stati raccolti campioni di sangue su cui è stata effettuata l’analisi del profilo lipidico. Negli ultimi anni, diversi studi hanno sottolineato la stretta associazione tra infiammazione a livello intestinale e peggioramento del quadro in pazienti con CKD. Il mantenimento di un ottimo stato del tratto gastrointestinale è fondamentale per assicurare lo stato di salute dell’ospite, contribuendo ai processi metabolici, fisiologici e immunologici. Le comunità batteriche instaurano un rapporto mutualistico con l’individuo che colonizzano, giocando un ruolo importante negli stati di salute e malattia. Un’anomala colonizzazione o cambiamenti nella composizione del microbiota intestinale, determina disbiosi, uno squilibrio associato a diverse condizioni patologiche come obesità, diabete di tipo II, malattia intestinale cronica, CVD e anche CKD. Il rapporto tra intestino e rene è bidirezionale, nei pazienti affetti da malattia renale cronica, la composizione del microbiota intestinale risulta essere modificata rispetto a quella del soggetto sano. Alti livelli di urea che si riversano facilmente nel tratto intestinale modificano il microambiente chimico con conseguente innalzamento del pH del colon che esercita una pressione selettiva a favore di specie ureasi-positive, responsabili della conversione dell’urea in ammoniaca. Lo strato protettivo di muco viene degradato e la permeabilità della barriera intestinale viene compromessa. In conseguenza di ciò si ha il passaggio di materiale batterico attraverso la mucosa e l’attivazione di un meccanismo infiammatorio. Nei pazienti con funzionalità renale compromessa, il rene perde progressivamente la capacità di eliminare sia le sostanze provenienti dal metabolismo umano, sia quelle della comunità microbica intestinale. Alcune di queste sostanze sono rappresentate dalle tossine uremiche, tra quelle di derivazione intestinale le principali e più studiate sono p-cresil solfato (PCS) e indossile solfato (IS). IS e p-CS, strettamente legate all’albumina sierica (Human Serum Albumin: HSA), non vengono eliminate facilmente ma rimangono nel torrente ematico. HSA è la più abbondante proteina sierica ed è la principale trasportatrice di composti esogeni ed endogeni, inclusi gli acidi grassi che sembrano rappresentare il maggior ligando endogeno della proteina. Multipli siti di legame vengono utilizzati per gli acidi grassi monoinsaturi (MonoUnsaturated Fatty Acids: MUFA) e PUFA. Acidi grassi e tossine uremiche competono quindi per gli stessi siti di legame sulla proteina. Il potenziale ruolo degli acidi grassi nel contrastare l’accumulo di tossine uremiche derivate dalla comunità batterica intestinale ne giustifica l’importanza della valutazione dei loro livelli ematici. Secondo scopo di questa tesi di dottorato è stato quello di valutare la possibile correlazione tra i livelli di acidi grassi circolanti e la composizione del microbiota intestinale in soggetti affetti da CKD. Sono stati arruolati nello studio 64 pazienti anziani con CKD non dializzati e 15 soggetti anziani con normale funzionalità renale. La composizione del microbiota intestinale è stata precedentemente caratterizzata attraverso l’impiego delle tecniche di elezione: PCR-DGGE e la PCR quantitativa (qPCR). In accordo con la letteratura scientifica, è stata evidenziata una riduzione di batteri saccarolitici e produttori di butirrato nei pazienti con CKD rispetto al gruppo di controllo. Il butirrato sembra giocare un ruolo cruciale nel mantenimento delle ottimali condizioni della barriera intestinale. Tenendo ciò in considerazione è stato deciso di approfondire lo studio e valutare l’associazione tra la comunità microbica intestinale e i livelli di acidi grassi basali in tali pazienti. Come risultato più importante ottenuto, è stata osservata una correlazione positiva statisticamente significativa tra la specie batterica Faecalibacterium Prausnitzii e i livelli totali di Omega-3 entrambi associati a proprietà antinfiammatorie. La presente tesi di dottorato evidenzia la necessità di sostenere ulteriori ricerche per supportare i risultati qui presentati. Studi futuri potrebbero essere utili per migliorare la comprensione del ruolo degli acidi grassi circolanti e i loro metaboliti sulla composizione del microbiota intestinale, sullo stato infiammatorio e sulla malattia renale cronica. / The aim of this thesis was to explore the possible associations between fatty acids (FA) profile and gut microbiota (gMb) with several conditions throughout the lifespan, from infancy to old age. In particular, we focused our attention on elderly subjects with Chronic Kidney Disease (CKD) and children with Acute Otitis Media (AOM). The terms “Chronic Kidney Disease” refers to several disorders with a progressive kidney function decline. International guidelines approved the definition of CKD as a condition with the presence of markers of kidney damage or with the estimated glomerular filtration rate (eGFR) less than 60 ml/min/1.73 m2 or both, for at least three months. End-stage renal disease is associated with a high cardiovascular disease (CVD) risk, the major cause of death in these patients. Chronic inflammation, oxidative stress, protein-energy wasting, disordered mineral metabolism, and deficiency of endogenous calcification inhibitors, known as non-traditional risks factor, take part in cardiovascular pathology in CKD. Inflammatory processes influence the physiological response to renal infection and injury but also participate in the development of potentially irreversible kidney damage with the production of various inflammatory molecular species, among whom eicosanoids and cytokines, from parental omega-6 long-chain polyunsaturated fatty acids (LCPUFA). Several studies focused their attention on the potential role of omega-3 (n-3) LCPUFA supplementation in subjects with CKD. Despite this, their effect on kidney damage is still not clear. However, it is widely agreed that a modified FA profile in CKD can determine a progression of the disease, inducing the inflammatory state. Moreover, high/normal n-3 LCPUFA levels decrease the risk of a decline of the disease. Omega-3 and omega-6 (n-6) LCPUFA concentrations and their ratios are tightly associated with renal health, because of their important roles in different pathways. Another aspect not very considered in the field of CKD is the role of circulating FA levels and their metabolites on the modulation of inflammation. The first aim of this study is to analyze the FA profile in elderly subjects with CKD. Blood samples have been collected from 57 subjects enrolled in the study, and FA analysis has been performed. During the last years, several studies underlined the strong relationship between intestinal inflammation and adverse outcomes in CKD. The health of gastrointestinal tract is fundamental to ensure the well being of the host contributing to its nutrition, metabolism, physiology, and immune function. The bacterial communities colonizing humans have been seen in terms of mutualistic symbiosis with their hosts, a mutually beneficial coexistence, playing an important role in health and disease. Abnormal colonization or changes in the gut microbial composition determine dysbiosis, a state associated with different illnesses, such as obesity, type 2 diabetes, inflammatory bowel disease, cardiovascular disease, and also chronic kidney disease. The relationship between gut and kidney is a bi-directional relation with a mutual influence. Chronic kidney disease influences gMB characteristics, especially through high levels of urea that easily spread in the intestinal fluid where bacterial urease enzymes degrade it, then it is hydrolyzed in ammonium hydroxide that increases fecal pH with a consequent alteration of intestinal cellular junctions. Besides, high levels of urea change intestinal microbiota composition damaging permeability of intestinal barrier and promoting proteolysis with production and absorption of uremic toxins, such as indoxyl sulfate (IS) and p-cresol sulfate (p-CS). These toxins induce an inflammatory process associated with CKD. Under physiologic conditions, the kidney through the urine eliminates these compounds, but CKD patients have a compromised renal clearance. Therefore, these solutes tend to accumulate in the organs. IS and p-CS are tightly bound to human serum albumin (HSA), the most abundant plasma protein in the bloodstream. HSA is recognized as the main means of transport for endogenous and exogenous compounds, including fatty acids that seem to be the main endogenous ligand of HSA, multiple binding sites are used for monounsaturated fatty acids (MUFA) and PUFA. Thus, free fatty acids and uremic toxins compete for the same binding sites on HSA. It is important to assess fatty acid (FA) levels in patients with CKD because of the potential role to contrast the accumulation of uremic toxins derived from the intestinal bacterial community. As a consequence of this bi-directional relation between gut and kidney and the possible involvement of some compounds as metabolites of FA in the inflammatory response, we investigate the correlation between circulating FA levels and the gMB composition in the same subjects with CKD, as the second aim of this thesis. 64 old CKD non-dialysis patients (eGFR 15-45 ml/min/1.73 m2) and 15 elderly subjects (>65 years) with normal renal function (eGFR >60 ml/min/1.73 m2, CKD-EPI) are enrolled. Bacterial composition was studied in a previous observational study by denaturating gel gradient electrophoresis (DGGE), high-throughput sequencing (16S ribosomal RNA), and quantitative real-time PCR (qPCR). This study described an increased abundance of some bacteria associated with pathological conditions. In agreement with the literature, the author found a reduced abundance of saccharolytic and butyrate-producing bacteria (Prevotella, Faecalibacterium prausnitzii, Roseburia) in CKD patients respect to the control group. Butyrate plays a crucial role in the maintenance of the gut barrier function. Taking that into account, we decided to investigate the correlation between gMB composition and FA profile in these subjects. The main result of the study was the significant positive correlation between Faecalibacterium prausnitzii and total n-3 levels, both associated with the antiinflammatory action. The present doctoral thesis underlines the need to perform further investigations in order to support evidence presented. Future studies may be useful to improve understanding of the effect of circulating fatty acids levels and their metabolites on gut microbial composition, inflammation process, and pathological conditions such as kidney disease. Our results showed that CKD patients with previous cardiovascular events had lower total and specific n-3 levels comparing with patients without them. Moreover, higher docosahexaenoic acid (DHA) levels and having had previous cardiovascular events seemed to have protective effects against further cardiovascular events. Moreover, we observed a significant reduction of the genera Roseburia and Faecalibacterium in CKD patients compared to C group and a significant lower abundance of F. prausnitzii and Roseburia spp. in CKD patients. Thus, our results seem in accordance with anti-inflammatory actions of total n-3, DHA, and saccharolytic and butyrateproducing bacteria. Many gMB changes seem to be related both to CKD and CVD. If the different gMB composition might play a causal role in cardiovascular events by an unbalanced production of some toxic substances, or if the gMB changes are merely a consequence of different dietary and lifestyle behaviours of these patients, it cannot be explained by the present study and all the yet available data. Further studies, possibly utilizing new high-throughput tools, will be required to understand the potential correlations between the gMB composition and other inflammation and oxidative stress markers in these patients. Other two studies have been performed during the doctoral course, to reach a better comprehension of fatty acids, gut microbial community and inflammatory states. A prospective pilot clinical study has been performed to to explore possible changes of gMB composition in children with AOM treated with amoxicillin with or without clavulanic acid. AOM is one of the most common bacterial infections in children and is normally treated with antibiotic therapies that lead to increasing antimicrobial resistance rates among otopathogens and may impair the correct development of the microbiota in early life. No significant differences were shown in the gMB composition of the overall cohort at different time intervals of the samples collection and in subjects treated with amoxicillin with or without clavulanic acid at different time intervals (T0, T1 and T2). A literature revision on lipids in infant formulae has been performed to better understanding quality and quality of dietary lipids because of their significant impact on health outcomes, especially when fat storing and/or absorption are limited (e.g., preterm birth and short bowel disease) or when fat byproducts may help to prevent some pathologies. The lipid composition of infant formulae varies according to the different fat sources used, and the potential biological effects are related to the variety of saturated and unsaturated FAs. Instead, ruminant-derived trans FAs and metabolites of n-3 LCPUFA with their anti-inflammatory properties can modulate immune function. Furthermore, dietary fats may influence the nutrient profile of formulae, improving the acceptance of these products and the compliance with dietary schedules. During the doctoral course, I spent a period abroad at Dell Pediatric Research Institute (DPRI), The University of Texas at Austin. In particular, I attended the laboratory of Doctor Brenna. I focused my research activity on a specific regulatory insertion-deletion polymorphism in the FADS gene cluster for better understanding its influence on PUFA and lipid profile.