By the year 2025, it is anticipated that over 300 million individuals world wide will have type 2 diabetes, with a projected increase from 84 to 288 million (170%) in developing countries and from 51 to 72 million (42%) in developed countries. Diabetes leads to a markedly increased risk of heart disease and renal failure and to expensive and debilitating retinopathy and neuropathy. Cognitive decline is also increased. As there is accumulating evidence of the beneficial effects of moderate carbohydrate, low fat dietary patterns compared to high carbohydrate diets, this thesis will focus on the effects of moderate carbohydrate high protein dietary patterns (total carbohydrate: protein: fat ratio of 40%:34%:26%) on glycemic control, risk factors for macrovascular disease and cognitive function. Information on two key areas in type 2 diabetes will be presented, 1. Acute effects of dietary patterns, moderately carbohydrate restricted and high in protein on glucose levels assessed using continuous glucose monitoring systems (CGMS) with verification of these results through a small repeat study. 2. Chronic effects of energy restricted dietary patterns, moderately carbohydrate restricted and high in protein on glucose levels, HbA1c, cognitive function, cardiovascular disease (CVD) risk markers and renal function. In the acute study, we recruited 23 subjects with type 2 diabetes. The participants were randomized to each of 4, 3-day interventions in a cross over design with a 4 day wash out period in which the carbohydrates were distributed differently at each meal; carbohydrates evenly distributed across the day, or carbohydrates loaded at breakfast, lunch or dinner. Glucose levels were continuously measured using CGMS. Outcomes were assessed by postprandial peak glucose (Gmax), time spent above 12 mmol/L (T>12) and total area under the glucose curve (AUC20). The intervention showed that an even distribution of carbohydrates did not optimise blood glucose control, whereas carbohydrates loaded at the lunch time meal provided the most favourable postprandial profile. To verify these results we conducted a repeat study. Six of the previous participants accepted the invitation to return and complete the even distribution arm of the study after a 20 week time lag. The intervention showed that although HbA1c, fasting blood glucose (FBG), AUC, exercise and ambient temperature remained constant there was a significant effect of change in sunlight hours on Gmax, suggesting an effect of sunlight. To assess the chronic effects of energy restricted dietary patterns on the determinants of HbA1c, cognitive function, CVD risk markers and renal function under conditions of weight loss, we recruited 82 participants with type 2 diabetes. These participants were randomised to one of two high protein energy restricted dietary patterns that differed in cholesterol content, for a 12 week period, in a parallel design. A sub group of these participants completed cognitive function testing with (n=34) or without (n=17) CGMS at baseline and at 8 weeks. After 8 weeks of the intervention the determinants of HbA1c under conditions of energy restriction were evaluated. The intervention showed the change in FBG accounted for most of the variance in change in HbA1c, but % energy reduction also contributed independently of FBG. Both energy restricted high protein diets equally improved glycemic control, particularly T>12, AUC, HbA1c and FBG. Fifty one participants completed cognitive testing to evaluate the effect of weight loss and blood glucose control on cognition. Cognitive function was not altered by time, diet, baseline lipid levels. Working memory was predicted by FBG. Short term memory was predicted by FBG, Gmax and AUC24. Sixty five participants completed 12 weeks of the intervention to assess CVD risk markers and renal function. Renal function was maintained and CV markers improved on both dietary patterns, with greatest improvement in HDL-C observed in the group consuming a high protein, energy restricted dietary pattern, high in dietary cholesterol. In conclusion, in the context of a high protein, carbohydrate restricted dietary pattern, cognitive function and renal function did not change, while glycemia and CV risk profiles improved with weight loss over the short term. Under conditions of energy balance diurnal glucose profiles were optimal when the carbohydrates were loaded in the lunch meal. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1342253 / Thesis (Ph.D.) - University of Adelaide, School of Molecular and Biomedical Science, 2008

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The renin-angiotensin-aldosterone system (RAAS) is very important during pregnancy, working mainly through the angiotensin II (Ang II) that can interfere with the activation of Mitogenic-Activated Protein Kinases (MAPK) cascade. In addition, pregnancy is associated with marked increase in renal hemodynamics that may also be affected by diabetes mellitus (DM). The objectives were to evaluate the effects of DM induced in Wistar rats during pregnancy and maintained in the postpartum period on the renal expression of PCNA (Proliferating Cell Nuclear Antigen), α-SMA (Smooth Muscle Actin) and p-p38 and p-JNK MAPK as well as in studies of renal function and systolic blood pressure (SBP) in Wistar rats. For this, there were two groups (G): non-pregnant and pregnant. In the first G, the rats were divided into: G1 (non-pregnant controls rats), females that received intraperitoneal injection (ipi) of 0.9% saline solution and G2 (non-pregnant diabetic rats), females that received ipi of alloxan (100 mg/kg) diluted in 0.9% saline solution. In the second G, pregnant rats were divided into: G3 (control mothers), mothers that received ipi of 0.9% saline solution and G4 (diabetic mothers), mothers that received ipi of alloxan (100 mg/kg) diluted in 0.9% saline solution. Blood glucose was observed 2 days after induction and the animals were considered diabetic if they had blood glucose higher than or equal to 150 mg/dL. About 50 days after delivery for G3 and G4 and corresponding time for G1 and G2, the rats were subjected to indirect measurement of SBP by plethysmography and the determination of glomerular filtration rate (GFR) by creatinine clearance. Then, the animals were anesthetized and their kidneys were removed for histological, immunohistochemical and morphometric studies. The results showed that G2 and G4 animals had blood glucose levels (mg/dL) and urinary volume (mL) higher than animals from G1 and G2, even several days after DM induction. The SBP (mmHg) were not different among the groups, but there was a tendency for reduction in GFR (mL/min) from G4 when compared to the other G. There was also a significant increase in kidney weight (right and left)/body weight ratio of G4 in relation to other G. Morphometric analysis showed a reduction of total renal sectional area from G4 and an increase in G3. The glomerular area and the capsular space did not differ between G studied, but there was an augment in glomerular tuft area in G3 and G4. Quantification of the percentage of cortical collagen showed that G2 and G4 presented higher distribution, while the identification and count of mast cells did not differ between G. Regarding PCNA immunoreactivity, G3 showed increased glomerular proliferating cells, while in G4 it was decreased. On the other hand, in the tubulointerstitial (TBI) compartment cell proliferation was higher in G4. Glomerular expression of α-SMA and TBI were increased in G4 compared to other G. Immunoreaction for glomerular p-p38 expression showed a pattern similar to PCNA, with a reduction of p-p38 in G4 relative to other G. The immunoreactivity of p-JNK was higher in both the glomeruli and TBI compartment in G4 compared to G1, G2 and G3. It is concluded that the DM induced during pregnancy and maintained in the postpartum period in Wistar rats resulted in significant structural changes and moderate functional impairmet in kidney, showing that pregnancy has an enhancing effect of renal damage induced by diabetes and these alterations may be related to changes in the expression of p-p38 and p-JNK MAPK. / O sistema renina-angiotensina-aldosterona (SRAA) é de suma importância durante a gestação, atuando principalmente através da angiotensina II (Ang II) que pode interferir na ativação da cascata de Mitogenic-Activated Protein Kinases (MAPK). Além disso, a gravidez está associada a aumentos marcantes na hemodinâmica renal, que também pode ser afetada pelo diabetes mellitus (DM). Os objetivos do trabalho foram avaliar os efeitos do DM induzido em ratas Wistar durante a gestação e mantido no período pós-parto na expressão renal de PCNA (Proliferating Cell Nuclear Antigen), α-SMA (Smooth Muscle Actin) e das MAPKs p-p38 e p-JNK, bem como nos estudos de função renal e pressão arterial sistólica (PAS) de ratas Wistar. Para isso, foram realizados dois grupos (G): não-gravídico e gravídico. No primeiro G, as ratas foram divididas em: G1 (controles sem gravidez), fêmeas que receberam injeção intraperitoneal (iip) de solução salina 0,9% e G2 (diabéticas sem gravidez), fêmeas que receberam iip de aloxana (100mg/kg) diluída em solução salina 0,9%. No segundo G, as ratas grávidas foram divididas em: G3 (mães controles), mães que receberam iip de solução salina 0,9% e G4 (mães diabéticas), mães que receberam iip de aloxana (100mg/kg) diluída em solução salina 0,9%. A glicemia foi verificada 2 dias após a indução e os animais foram considerados diabéticos se apresentassem glicemia maior ou igual a 150mg/dL. Cerca de 50 dias após o parto para G3 e G4 e em tempo correspondente para G1 e G2, as ratas foram submetidas à aferição indireta de PAS por pletismografia, bem como à determinação da Taxa de Filtração Glomerular (TFG) pelo clearance de creatinina. Em seguida, os animais foram anestesiados e tiveram seus rins retirados para as análises histológica, morfométrica e imunohistoquímica. Os resultados mostraram que os animais de G2 e G4 apresentaram glicemia (mg/dL) e volume urinário (mL) maiores que os animais de G1 e G2 vários dias pós-indução do DM do tipo 1. Os valores de PAS (mmHg) não foram diferentes entre os grupos analisados, mas constatou-se uma tendência à redução da TFG (mL/min) de G4 quando comparado aos demais G. Observou-se também um aumento significativo da relação peso rim/peso do corpo (direito e esquerdo) dos animais de G4 em relação aos demais G. As análises morfométricas evidenciaram redução da área de secção renal total de G4 e aumento em G3. A área glomerular e o espaço capsular não diferiram entre os G estudados, mas houve aumento da área de tufo glomerular em G3 e G4. A quantificação da porcentagem de colágeno cortical mostrou que G2 e G4 tiveram maior expressão, ao passo que a identificação e contagem de mastócitos não diferiram entre os G. Quanto à imunorreação para PCNA, o G3 apresentou aumento de células PCNA+ glomerulares, ao passo que em G4 diminuiu. Por outro lado, no compartimento tubulointersticial (TBI) a proliferação celular foi maior em G4. Já a expressão de α-SMA glomerular e TBI foram aumentadas em G4 em comparação aos demais G. A marcação para p-p38 glomerular apresentou um padrão de expressão semelhante ao PCNA, com redução de células p-p38+ em G4 em relação aos outros G. A imunorreação para p-JNK mostrou-se elevada tanto nos glomérulos quanto no compartimento TBI de G4 em relação a G1, G2 e G3. Conclui-se que o DM induzido durante a gestação e mantido no período pós-parto em ratas Wistar resultou em alterações estruturais significativas e funcionais renais moderadas, evidenciando que a gravidez possui um efeito intensificador dos danos renais induzidos pelo diabetes e que estas alterações podem estar relacionadas com as mudanças na expressão das MAPK p-p38 e p-JNK. / Mestre em Biologia Celular e Estrutural Aplicadas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Hypothyroidism, in both its clinical and subclinical manifestations, is a highly prevalent endocrinopathy among pregnant women. Given the high fetal dependency on maternal thyroid hormones (TH), a shortage of these hormones can be related to long-term effects on organisms, as TH have ubiquitous actions. Particularly, effects of gestational hypothyroidism on kidneys could cause an elevation of blood pressure later in life. An indicator of fetal programming can be the somatic growth of an organism, which, in rats, is hard to be modelled due to persistent growth post-puberty. With this purpose, use of body weight in analysis of variance (ANOVA), taking animal age as repeated measures factor (RMF), can cause nonsphericity in this statistical model. A goal was to evaluate, on adult offspring, the influence of maternal TH deficiency on renal function and on somatic growth, as well the relationship between renal function and systolic arterial pressure (SAP). Additionally, we sought an adequate method of analysis for body weight curves of rats and investigated consequences of using ANOVA as usual for this purpose. Female Wistar rats were divided in three main experimental groups: control euthyroid; hypothyroid by addition of methimazole (0.02%) in drinking water; and euthyroid by hormonal replacement, receiving both methimazole (0.02%) and levothyroxine (T4, 100 μg/L) in drinking water. Treatment was given between gestational days (GD) 9 and 21. On offspring, either nephrectomy or sham-surgery was done at post-coitus day (PCD) 26. Blood pressure was evaluated at around postnatal day (PND) 90, through a tail-plethysmograph. Blood and urine were collected at around PND 110 to evaluate creatinine clearance. To evaluate somatic growth, animals were weighed once to thrice weekly after weaning. Analysis of weight data was done by fitting a modified tetraparametric sigmoid curve to weight data and reducing its parameters by principal component analysis, whereby two principal components were able to retain 92% of original variance of parameters. Experimental gestational hypothyroidism (EGH) had no effect on SAP (p* > 0,200), except through its interaction with estrous cycle. There was a third order interaction concerning estrous phase, nephrectomy and T4 replacement, where the luteal SAP drop was intensified (p* = 0,039). Body weight at PCD 26 did not show any correlation with SAP (p* = 0,716). There was no correlation between SAP and creatinine clearance (p* = 0,803). EGH had no significant effect on creatinine clearance nor on weight curve principal components (p* > 0,200). Thus, given current approach, it was not possible to detect any EGH effect on SAP, neither on renal function, nor on somatic growth. Analysis of body weight curves allowed a precise definition of the beginning of adulthood of animals regarding their somatic growth and elaboration of a simplified model of somatic growth of rats. Also, it was possible to establish an empirical maximum for the coefficient of sphericity of studies which use repeated measures ANOVA to model continuous variables which have been discretized and used as RMF; this can be useful for both experimental planning and previous results reevaluation. / O hipotireoidismo, em suas formas clínica e subclínica, é endocrinopatia de prevalência significativa em mulheres gestantes. Dada a dependência fetal de hormônios tireoideanos (HT) maternos, a sua carência pode estar relacionada a efeitos duradouros no organismo, tendo-se em vista a ação ubíqua dos HT. Em particular, efeitos do hipotireoidismo gestacional no desenvolvimento renal têm o potencial de levar a uma elevação da pressão arterial (PA) na vida adulta. Um indicador da programação fetal pode ser o crescimento somático do organismo, que, em ratos, é difícil de ser modelado devido ao crescimento persistente pós-puberal. Nesse sentido, o uso do peso corporal em análises de variância (ANOVA), tomando-se como fator de medidas repetidas (FMR) a idade do animal, é capaz de violar a premissa de esfericidade do modelo estatístico. Um dos objetivos do estudo foi avaliar, na vida adulta da prole de ratas, a influência da carência dos HT maternos na função renal e no crescimento somático, bem como a relação da função renal com a PA sistólica (PAS). Além disso, buscou-se caracterizar um método adequado de análise de curvas de peso corporal de ratos e investigar as consequências do uso de ANOVA em sua forma tradicional para esse propósito. Ratas Wistar prenhas foram divididas em três grupos experimentais principais: eutireoideanas controle; hipotireoideanas por adição de metimazol (0,02%) na água de beber; e eutireoideanas por reposição hormonal, recebendo tanto metimazol (0,02%) quanto levotiroxina (T4, 100 μg/L) na água de beber. Os tratamentos foram realizados do 9º ao 21º dia de gestação. Com as proles das ratas, foram feitas: cirurgias de nefrectomia ou pseudocirurgia equivalente no 26º dia pós-cópula (DPC); medidas de PAS com cerca de 90 dias pós-natal (DPN) através de um pletismógrafo de cauda; e, coletas de sangue e urina para avaliação do clearance de creatinina com cerca de 110 DPN. Para a avaliação do crescimento somático, os animais foram pesados entre 1 e 3 vezes por semana após o desmame. A análise de curvas de peso corporal foi feita por ajuste de sigmoide de quatro parâmetros, reduzidos a dois componentes principais, mantendo-se 92% da variância original dos parâmetros. O hipotireoidismo gestacional experimental (HGE) não teve efeito significativo sobre a PAS (p* > 0,200), exceto em sua interação com o ciclo estral: houve interação do ciclo estral com a reposição de T4 da mãe e nefrectomia, em que a queda de PAS da fase lútea foi acentuada (p* = 0,039). O peso corporal no 26º DPC não apresentou correlação com a PAS (p* = 0,716). Também não houve correlação entre a PAS e o clearance de creatinina (p* = 0,803). O HGE não teve efeito significativo sobre o clearance de creatinina nem sobre os componentes principais das curvas de peso corporal (p* > 0,200). Portanto, não foi possível verificar, com a abordagem utilizada, qualquer efeito do HGE sobre a PA da prole adulta, nem sobre a função renal e tampouco sobre o crescimento somático. A análise do peso corporal permitiu uma definição precisa do início da vida adulta dos animais quanto ao seu crescimento somático e a elaboração de um modelo conceitual simplificado do crescimento somático de ratos. Além disso, foi possível estabelecer um máximo empírico para o coeficiente de esfericidade de estudos que usam ANOVA de medidas repetidas para modelar variáveis contínuas que tenham sido discretizadas e usadas como FMR; isso pode ser útil tanto no planejamento experimental quanto na reavaliação de achados anteriores. / São Cristóvão, SE

Made available in DSpace on 2016-01-26T18:55:31Z (GMT). No. of bitstreams: 1 DISSERTACAO.pdf: 506827 bytes, checksum: 85183faa6a63e1b561cacb568f6cf77e (MD5) Previous issue date: 2011-02-24 / The anti-inflammatory nonsteroidal compounds have extremely widespread use in the therapy of small animals, due to their anti-inflammatory and analgesic properties. However, the use of these drugs can produce changes in kidney function. The present study was conducted to assess kidney function in healthy dogs undergoing therapy with anti-inflammatory nonsteroidal compounds. Thirty mongrel dogs, adults, males and females, clinically healthy, were divided randomly into 5 groups (G) of six animals each receiving the following therapies: Gceto ketoprofen, a 2 mg/kg dose (VO), every 24 hours, during 10 days; Gnime nimesulide, 5 mg/Kg, VO, every 24 hours, during 10 days; Gmelo- meloxican, 0.2 mg/Kg on the first day, followed by 0.1 mg/Kg, VO, every 24 hours, 7 days; Geto etodolac, 15 mg/Kg, VO, every 24 hours, 7 days; Gcele- celecoxibe, 5 mg/Kg, VO, every 12 hours, for 20 days. The physical examination and renal function (urinalysis, urinary GGT, creatinine and sodium, serum urea, creatinine, potassium and sodium, and endogenous creatinine clearance) were assessed prior to the treatment, on the 5th and 10th days (T0, T5 and T10) into the treatment in all groups, and also on the 20th day (T20) into the treatment in Gcele. Few changes were observed in urinalysis parameters, only with significant increase in the presence of renal cells in the urine in T5, in the nimesulide group. There was a significant reduction in sodium elimination in the animals urine in the nimesulide group. The clearance values showed significant decrease in the celecoxibe group in T20, in relation to T5. The enzyme GGT urinary showed no variation among groups or moments. Values of sodium, potassium, urea and creatinine serum remained within normal ranges in all times, in the different groups. In conclusion, minimal changes of renal function occur 10 days after therapy on set with NSAIDS nimesulide, after 10 days with ketoprofen, on the 20th days of therapy with celecoxibe in health dogs. / Os anti-inflamatórios não-esteroidais têm uso extremamente difundido na clínica de pequenos animais, devido às suas propriedades analgésicas e anti-inflamatórias. Entretanto, o uso desses fármacos pode produzir alterações da função renal. O presente estudo teve como objetivo avaliar a função renal de cães saudáveis, submetidos à terapia com anti-inflamatórios não-esteroidais não seletivos, COX-2 preferenciais e COX-2 seletivos. Foram utilizados 30 cães, sem raça definida, adultos, machos e fêmeas, clinicamente sadios, divididos aleatoriamente em 5 grupos (G) de 6 animais cada, que receberam as seguintes terapias: Gceto cetoprofeno, na dose de 2 mg/Kg, por via oral (VO), a cada 24 horas, durante 10 dias; Gnime nimesulida, 5 mg/Kg, VO, a cada 24 horas, durante 10 dias; Gmelo - meloxican, 0,2 mg/Kg no primeiro dia, seguido por 0,1 mg/Kg, VO, a cada 24 horas, por 10 dias; Geto etodolaco, 15 mg/Kg, VO, a cada 24 horas, 10 dias; Gcele - celecoxibe, 5 mg/Kg, VO, a cada 12 horas, por 20 dias. O exame físico e a função renal (urinálise; GGT, creatinina e sódio urinários; uréia, creatinina, sódio e potássio séricos; e clearance endógeno de creatinina) foram avaliados antes, aos 5, 10 dias (T0, T5 e T10) de tratamento em todos os grupos, e também aos 20 dias (T20) de tratamento no Gcele. Poucas alterações foram observadas na urinálise, apenas com aumento significativo da presença de células renais na urina no T5 e T10 em relação ao T0, no grupo nimesulida. Houve redução significativa da eliminação de sódio na urina, nos animais do grupo nimesulida, no T5. Os valores de clearance foram os mais baixos no grupo Cetoprofeno no T10, e revelaram diminuição significativa no grupo Celecoxibe no T20, em relação ao T5. A enzima GGT urinária não apresentou variação entre grupos ou momentos. Valores de sódio, potássio, uréia e creatinina séricos mantiveram-se dentro da normalidade em todos os momentos nos diferentes grupos. Conclui-se que, em cães hígidos, alterações mínimas da função renal ocorrem aos cinco dias de terapia com o AINE nimesulida, aos dez dias com cetoprofeno, e aos vinte dias de terapia com celecoxibe.

Made available in DSpace on 2016-07-18T17:53:08Z (GMT). No. of bitstreams: 1 DISSERTACAO.pdf: 506827 bytes, checksum: 85183faa6a63e1b561cacb568f6cf77e (MD5) Previous issue date: 2011-02-24 / The anti-inflammatory nonsteroidal compounds have extremely widespread use in the therapy of small animals, due to their anti-inflammatory and analgesic properties. However, the use of these drugs can produce changes in kidney function. The present study was conducted to assess kidney function in healthy dogs undergoing therapy with anti-inflammatory nonsteroidal compounds. Thirty mongrel dogs, adults, males and females, clinically healthy, were divided randomly into 5 groups (G) of six animals each receiving the following therapies: Gceto ketoprofen, a 2 mg/kg dose (VO), every 24 hours, during 10 days; Gnime nimesulide, 5 mg/Kg, VO, every 24 hours, during 10 days; Gmelo- meloxican, 0.2 mg/Kg on the first day, followed by 0.1 mg/Kg, VO, every 24 hours, 7 days; Geto etodolac, 15 mg/Kg, VO, every 24 hours, 7 days; Gcele- celecoxibe, 5 mg/Kg, VO, every 12 hours, for 20 days. The physical examination and renal function (urinalysis, urinary GGT, creatinine and sodium, serum urea, creatinine, potassium and sodium, and endogenous creatinine clearance) were assessed prior to the treatment, on the 5th and 10th days (T0, T5 and T10) into the treatment in all groups, and also on the 20th day (T20) into the treatment in Gcele. Few changes were observed in urinalysis parameters, only with significant increase in the presence of renal cells in the urine in T5, in the nimesulide group. There was a significant reduction in sodium elimination in the animals urine in the nimesulide group. The clearance values showed significant decrease in the celecoxibe group in T20, in relation to T5. The enzyme GGT urinary showed no variation among groups or moments. Values of sodium, potassium, urea and creatinine serum remained within normal ranges in all times, in the different groups. In conclusion, minimal changes of renal function occur 10 days after therapy on set with NSAIDS nimesulide, after 10 days with ketoprofen, on the 20th days of therapy with celecoxibe in health dogs. / Os anti-inflamatórios não-esteroidais têm uso extremamente difundido na clínica de pequenos animais, devido às suas propriedades analgésicas e anti-inflamatórias. Entretanto, o uso desses fármacos pode produzir alterações da função renal. O presente estudo teve como objetivo avaliar a função renal de cães saudáveis, submetidos à terapia com anti-inflamatórios não-esteroidais não seletivos, COX-2 preferenciais e COX-2 seletivos. Foram utilizados 30 cães, sem raça definida, adultos, machos e fêmeas, clinicamente sadios, divididos aleatoriamente em 5 grupos (G) de 6 animais cada, que receberam as seguintes terapias: Gceto cetoprofeno, na dose de 2 mg/Kg, por via oral (VO), a cada 24 horas, durante 10 dias; Gnime nimesulida, 5 mg/Kg, VO, a cada 24 horas, durante 10 dias; Gmelo - meloxican, 0,2 mg/Kg no primeiro dia, seguido por 0,1 mg/Kg, VO, a cada 24 horas, por 10 dias; Geto etodolaco, 15 mg/Kg, VO, a cada 24 horas, 10 dias; Gcele - celecoxibe, 5 mg/Kg, VO, a cada 12 horas, por 20 dias. O exame físico e a função renal (urinálise; GGT, creatinina e sódio urinários; uréia, creatinina, sódio e potássio séricos; e clearance endógeno de creatinina) foram avaliados antes, aos 5, 10 dias (T0, T5 e T10) de tratamento em todos os grupos, e também aos 20 dias (T20) de tratamento no Gcele. Poucas alterações foram observadas na urinálise, apenas com aumento significativo da presença de células renais na urina no T5 e T10 em relação ao T0, no grupo nimesulida. Houve redução significativa da eliminação de sódio na urina, nos animais do grupo nimesulida, no T5. Os valores de clearance foram os mais baixos no grupo Cetoprofeno no T10, e revelaram diminuição significativa no grupo Celecoxibe no T20, em relação ao T5. A enzima GGT urinária não apresentou variação entre grupos ou momentos. Valores de sódio, potássio, uréia e creatinina séricos mantiveram-se dentro da normalidade em todos os momentos nos diferentes grupos. Conclui-se que, em cães hígidos, alterações mínimas da função renal ocorrem aos cinco dias de terapia com o AINE nimesulida, aos dez dias com cetoprofeno, e aos vinte dias de terapia com celecoxibe.

La polyneuropathie sensitivomotrice héréditaire (PNSMH) est une maladie rare qui entraîne un ralentissement du développement moteur et mental, une déficience sensitivomotrice et des syndromes neuropsychiatriques, et qui s’accompagne souvent d’une agénésie du corps calleux. Par ailleurs, plusieurs évaluations rapportent une petite stature ou une masse corporelle anormalement basse chez les patients. La PNSMH est causée par des mutations perte de fonction du cotransporteur K⁺-Cl⁻ de type 3 (KCC3). Des évaluations cliniques détaillées et la caractérisation de souris inactivées pour Kcc3 (Kcc3ᴷᴼ) ont permis d’établir qu’un défaut d’export K⁺-Cl⁻ cause les atteintes neurologiques anatomiques et fonctionnelles dans la maladie. Chez les souris Kcc3ᴷᴼ, des manifestations extraneurologiques ont également été relevées : masse corporelle réduite, pression artérielle (PA) élevée, polydipsie et polyurie. Puisque la physiopathologie des désordres extraneurologiques découlant de la perte de fonction de KCC3 reste incomplètement décrite, mes travaux avaient pour objectif d’en comprendre les mécanismes sous-jacents en utilisant un modèle Kcc3ᴷᴼ. Une caractérisation initiale de notre lignée de souris Kcc3ᴷᴼ constitutive et systémique a montré des anomalies vasculaires et cardiaques accompagnant une élévation de PA diastolique. Cette lignée affichait également une polydipsie et une polyurie isoosmotique, de même qu’une réduction de masse corporelle et d’adiposité sans réduction d’apport alimentaire. Une caractérisation métabolique détaillée de notre modèle a ensuite permis de révéler des réductions de masse grasse et de masse maigre. Cette minceur résulte sûrement en partie des augmentations d’activité locomotrice et de dépense énergétique mesurées. Une nette amélioration de la tolérance au glucose a aussi été trouvée, ainsi que des concentrations réduites de triacylglycérols plasmatiques. Enfin, nous avons noté que notre modèle est résistant à l’obésité induite par une diète hyperlipidique et affiche une élévation concomitante de l’expression d’enzymes lipogéniques et lipolytiques dans le gras viscéral, engendrant potentiellement une dissipation calorique. En revisitant la fonction cardiovasculaire dans notre modèle par des méthodes de pointe, nous n’avons pas observé de changement de PA ni de différence de réactivité artériolaire en conditions basales, mais nous avons noté une élévation de distensibilité artériolaire passive. Chez notre modèle, nous n’avons pas non plus remarqué de sensibilité particulière de la PA au sel alimentaire, mais une excrétion urinaire fortement accrue de solutés sous diète hypersodée ainsi qu’une préférence marquée pour le sel. Ces observations sont compatibles avec un défaut de réabsorption hydrosodée par le rein pouvant d’ailleurs prévenir les élévations de PA. En somme, nos travaux ont permis de mieux comprendre les atteintes cardiométaboliques qui accompagnent le tableau neurologique d’un modèle murin de PNSMH. Nous avons notamment relevé des bénéfices inattendus dans le métabolisme glucidique et lipidique suivant l’inactivation de Kcc3. Nous soupçonnons également que l’absence de KCC3 dans le rein engendre une fuite ionique urinaire s’accentuant sous diète hypersodée et pouvant influencer la PA en limitant l’expansion volémique. Nos observations d’anomalies pléiotropiques liées à l’inactivation de Kcc3 font de ce gène une nouvelle cible pharmacologique potentielle et justifient la nécessité d’étudier l’anatomophysiologie cardiométabolique des patients atteints de PNSMH de façon plus approfondie. / Hereditary motor and sensory neuropathy (HMSN) is a rare disease that leads to delayed motor and mental development, loss of sensory and motor function and neuropsychiatric syndromes, and that is often accompanied by partial or complete agenesis of the corpus callosum. Additionally, several cases of short stature or low body weight have been reported in patients. HMSN is caused by loss-of-function mutations in K⁺-Cl⁻ cotransporter type 3 (KCC3). Detailed clinical reports and characterizations of mice inactivated for Kcc3 (Kcc3ᴷᴼ) have allowed to establish that defective K⁺-Cl⁻ export causes the anatomical and functional neurologic impairments in the disease. In Kcc3ᴷᴼ mice, extra-neurological abnormalities have also been noted: lower body weight, high blood pressure (BP), polydipsia and polyuria. Because the pathophysiology of extra-neurological traits arising from KCC3 loss of function remains incompletely described, my work aimed at understanding the mechanisms at play using a Kcc3ᴷᴼ model. An initial characterization of a constitutive and systemic Kcc3ᴷᴼ mouse line showed vascular and cardiac abnormalities along with a rise in diastolic BP. This model also showed polydipsia and iso-osmolar polyuria along with reduced body weight and adiposity but no decrease in food intake. A detailed metabolic characterization of our model further revealed reductions in fat and lean body masses. This leanness results certainly in part from increased locomotor activity and energy expenditure as measured. A marked improvement in glucose tolerance was also found in addition to lower plasmatic triglyceride concentrations. Lastly, we also demonstrated that our model is resistant to high-fat-diet-induced obesity and shows concomitant increase in expression of both lipogenic and lipolytic enzymes in visceral fat, thereby potentially generating caloric dissipation. When revisiting the cardiovascular function of our model with cutting-edge methods, we measured normal BP and arteriolar reactivity in baseline conditions. However, we noted an increase in passive arteriolar distensibility. In our model, we did not notice sensitivity of BP to dietary salt but found a marked increase in urinary solute excretion under high-salt diet and a strong preference for salt. These observations are consistent with a defect in hydromineral reabsorption by the nephron that may prevent BP from rising. In short, our work allowed to better understand the cardiometabolic characteristics that accompany the neurologic portrait of an HMSN mouse model. In particular, we noted unexpected benefits in carbohydrate and lipid metabolism upon Kcc3 inactivation. We also suspect that KCC3 ablation in the kidney leads to urinary hydromineral wasting that can be more salient under dietary salt loading and can influence BP by blunting extracellular volume expansion. The pleiotropic abnormalities arising from Kcc3 inactivation identify this gene as a new potential pharmacological target and argue for improving efforts at describing the cardiometabolic features of patients with HMSN.

Trotz Fortschritte in den letzten Dekaden ist das Langzeitüberleben von Nierentransplantaten unzureichend. Die Personalisierung der Behandlung kann dabei zu erheblichen Verbesserungen führen. Vor diesem Hintergrund wurde eine Kohorte von 587 Patienten im ersten Jahr nach der Transplantation untersucht und ein breites Spektrum von Markern zur langfristigen Prognose etabliert. In dieser Dissertation beschreibe ich in vier Manuskripten und zwei Kapiteln meine Arbeit zur personalisierten Transplantationsmedizin. Der klinische Verlauf von Patienten nach Nierentransplantation wurde untersucht. Die wichtigen Komplikationen standen im Vordergrund: Virusreaktivierungen – insbesondere die BK- und Cytomegalieviren – und akute Abstoßung. Folgende Analysen wurden durchgeführt: (i) Systematische Analyse der Assoziationen zwischen Virusreaktivierungen und deren Einfluss auf das Transplantationsergebnis; (ii) Bewertung der Auswirkungen antiviraler Behandlungsstrategien auf die Transplantationsergebnisse; (iii) Entwicklung eines Tools zur Prätransplantations-Risikoeinschätzung der Abstoßung und (iv) Erstellung eines mathematischen Modells für die personalisierte Charakterisierung der Immunantwort gegen das BK-Virus. Zusammengenommen haben die vier Studien das Potenzial, (i) die Patientenversorgung zu verbessern, (ii) die Überwachung von Virusreaktivierungen zu optimieren, (iii) Präventionsstrategien gegen virale Reaktivierungen zu stratifizieren, (iv) die Behandlung der Patienten an das individuelle Risiko akuter Abstoßung anzupassen, und (v) zur Personalisierung der Immuntherapie beizutragen. Die Studien zeigen, wie das große Datenvolumen einer klinischen Studie zur Weiterentwicklung der personalisierten Medizin unter Einsatz effektiver Strategien für Datenmanagement, Analyse und Interpretation genutzt werden kann. Es ist zu erwarten, dass diese Ergebnisse die klinische Praxis beeinflussen und so das langfristige Überleben und die Lebensqualität der Patienten verbessern. / In spite of the developments in the last decades, long-term graft survival rates in kidney transplantation are still poor: Personalization of treatment can thereby lead to a drastic improvement in long-term outcomes. With this goal, a cohort of 587 patients was characterized for a wide range of markers during the first post-transplantation year to assess their long-term prognosis. Here, I describe along four manuscripts and two chapters my work on personalized medicine for renal transplantation. In detail, we have studied the clinical evolution of patients with emphasis on two most relevant complications: viral reactivations – particularly those of BK virus and cytomegalovirus – and acute rejection. We have analysed in depth these phenomena by (i) exhaustively analysing the associations between different viral reactivations and their influence on transplantation outcome, (ii) evaluating the effects of antiviral treatment strategies on viral reactivation and other transplantation outcomes with emphasis on sex-associated differences, (iii) developing a tool for the pre-transplantation risk assessment of acute cellular rejection, and (iv) creating a mathematical model for the personalized characterization of the immune response against the BK virus under immunosuppression. Taken together, these studies have the potential of improving patient care, optimizing monitoring of viral reactivations, stratifying antiviral prevention strategies, tailoring immunosuppression and monitoring to the individual risk of acute rejection, and contributing to personalization of immunotherapy. They demonstrate how the large volume of data obtained within a clinical study can be employed to further the development of personalized medicine, employing effective data management, analysis and interpretation strategies. We expect these results to eventually inform clinical practice, thereby improving long-term survival and quality of life after kidney transplantation.