O processo inflamatório subclinico encontra-se associado à prevenção e controle de um agrupamento de fatores de risco (FR) nutricionais, entre eles, dislipidemia e aumento de depósito de gordura visceral. Indicadores nutricionais, inflamatórios e metabólicos parecem estar associados com o estilo de vida. A semente de linhaça tem sido reconhecida como um alimento rico em fibras e -3, entretanto, um novo constituinte de sua composição nutricional tem merecido atenção, pelo seu papel antiinflamatório e antioxidante. Este componente é chamado de lignanas, um polímero complexo e o principal constituinte não-carbohidrato de plantas vasculares. Está ligado a fibras de celulose, e é responsável por reforçar a estrutura das paredes celulares, o que previne o colapso das mesmas. Lignanas, em contato com a microflora intestinal humana transformam-se em enterolignanas, especialmente, enterodiol e enterolactona. O presente trabalho tem por hipótese que o teor de lignanas dietético pode interferir no perfil metabólico, e alterar fatores de riscos envolvidos no estado nutricional, e consequentemente na saúde. O conhecimento de que diferentes características na composição nutricional de macronutrientes da dieta poderiam modificar o perfil inflamatório, independentemente, da presença das enterolignanas provenientes da semente de linhaça também constituíram o objetivo deste estudo. Por 42 dias, foram avaliados 52 funcionários, pertencentes ao sexo masculino, com idade média de 37± 9 anos, de uma indústria de grande porte, na cidade de Itu-SP. Os voluntários foram divididos em 4 grupos de pesquisa, sendo, um grupo controle, e três grupos com dietas isocalóricas e diferentes proporções no % de carboidratos (CH), e acréscimo de semente de linhaça em pó ou arroz cru triturados (protocolo duplo cego). Foi preenchida ficha de coleta de informações sobre dados pessoais e conhecimento de fatores de risco (hipertensão, dislipidemia e diabetes), comportamentos de risco (tabagismo e sedentarismo) e antecedentes familiares. Foi também realizada avaliação clinico - laboratorial, no qual se obteve o registro de medidas antropométricas, medida da pressão arterial e coleta de sangue venoso em jejum de 12 h para avaliação de indicadores bioquímicos referentes à FR cardiovascular, tais como, colesterol total e frações (LDL-c e HDL-c), triglicérides, glicemia, insulina, Homa-beta e Homa-IR, ácido úrico, bem como, para avaliação de indicadores inflamatórios (Proteína C Reativa (PCR), Fator de Necrose Tumoral (TNF-) e Isoprostane Sérico), hormonais (Leptina e Adiponectina) e nutricionais (Enterodiol e Enterolactona séricas e urinárias). Observou-se que para redução significativa das medidas antropométricas estudadas e indicador de estresse oxidativo não houve diferenças entre os grupos que receberam intervenção dietética. Entretanto, para a melhora do perfil bioquímico, inflamatório, hormonal e nutricional, diferentes respostas foram encontradas. Os grupos que receberam dietas com redução de CH total (32% e 35%) mostraram benefícios, no que se refere ao perfil bioquímico, especialmente, colesterol total, LDL-c e ácido úrico, como também, para o perfil hormonal, referente aos níveis de adiponectina (p <0,05). Com relação aos níveis de PCR e TNF-, apenas os grupos que tiveram acréscimo de semente de linhaça na dieta apresentaram redução significativa (p<0,05). Para os níveis de triglicérides, somente o grupo com adição de semente de linhaça e 32 % de CH total apresentou diminuição significativa (p<0,05). Foi observado que com 32 % de CH total ingerido e adição de um alimento rico em lignanas constituiu-se uma estratégia nutricional relevante, para prevenção primária de fatores de risco metabólicos e controle da inflamação subclinica, o que pode contribuir na redução da morbi-mortalidade a eles associada. / The control of subclinical inflammatory process is associated with the prevention nutritional RF (risk factor), such as dislipidemia and the increase of visceral fat deposition. Nutritional, inflammatory and metabolic indicators seem also to related to life style. The linseed has been recognized as rich in fibers and -3. However, a new component in its nutritional composition has deserved the attention for its anti- inflammatory and antioxidant roles. This component is called lignans, a complex polymer and the main non-carbohydrate constituent of vascular plants. It is binded to cellulose fibers and is responsible for reinforcing cell walls structure, preventing them from collapsing. Plant lignans, in contact with the human intestinal flora, become enterolignans, specially enterodiol and enterolactone. This present work hypothetically that the amount of dietetic plant lignans interfere in the metabolic profile, altering the risk factors involved in the nutritional health state and consequently, the welfare state. Therefore, the objective of this study is to know if different diet nutritional composition characteristics can change the inflammatory profile, independently of the presence of enterolignans from the linseed. For 42 days, 52 male volunteers, average 37±9 years old, from a industrial city of Itu-SP, were evaluated. The volunteers were divided into 4 research groups; one control group and 3 groups on isocaloric diets with different proportion of carbohydrate (CH) and the addition of powdered linseed or ground raw rice (a double blind protocol). Personal data, RF (hypertension, dislipidemia and diabetes), habits (smoking and sedentary) and family antecedents were collected. A nutritional-laboratorial evaluation was performed in order to get anthropometric data; blood pressure checked and blood samples (after 12 hours fast) for total cholesterol and fractions (LDL-c and HDL-c ), triglycerides, glycemia, insulin, Homa-beta and Homa-IR, uric acid, inflammatory indicators (Reactive-C Protein (PCR) , Tumoral Necrosis Factor (TNF-a) and Seric Isoprostane ), hormonal (Leptin and Adiponectin), enterodiol and enterolactone seric and urinary enterolignins. It was observed a reduction of the studied anthropometric measures and for the oxidative stress indicators. A significant change occurred in the anthropometric measurements and the oxidative stress marker evaluated for all groups, but no difference among them was noted. On the other hand, biochemical, inflammatory, hormonal and nutritional profile significant differences among groups was observed. The groups that received diets with the reduction of the total CH (32% and 35% ) showed improvements in the biochemical profile, specially in the total cholesterol, LDL-c and uric acid, as well as the hormonal profile, in the levels of adiponectin (p< 0,05) . The levels of PCR and TNF-a, only the groups that had the linseed, showed a reduction (p< 0,05). For the triglycerides levels, only the group with the addition of linseed and 32% of total CH showed a decrease. It was observed with 32% of CH and the addition of food lignans constitute a nutritional relevant strategy for the primary prevention of metabolic risk factors and control of subclinical inflammation, contributing to the reduction of the associated morbi-mortality.

La ménopause est un facteur de risques associé au développement des maladies métaboliques. Du fait de ce déficit hormonal, les femmes ménopausées pourraient présenter des risques particuliers une fois exposées à des agents chimiques capables de perturber le système endocrinien. Pour en savoir plus, nous avons mis au point un modèle de souris ovariectomisées supplémentées ou non en œstradiol (E2), et exposées de manière chronique à un mélange de polluants (TCDD, PCB153, DEHP et BPA) ajoutés à une alimentation obésogène. Chaque polluant est utilisé à une dose équivalente à la Dose Journalière Tolérable, et l’exposition recouvre différents stades de développement, incluant ou non la période maternelle (gestation + lactation). Nos résultats indiquent qu’en condition d’obésité induite par le régime, l’ovariectomie accentue la prise de poids, l’intolérance au glucose et l’insulino-résistance chez les souris femelles adultes. Ce phénotype est associé à une diminution de la signalisation œstrogénique dans le foie et une dérégulation de l’adipogenèse et de l’inflammation dans le tissu adipeux. De manière intéressante, le mélange de polluants réduit l’impact délétère de l’ovariectomie sur le phénotype métabolique, possiblement par une amélioration de la signalisation œstrogénique dans le foie et le tissu adipeux. Nos résultats suggèrent fortement que ces effets dépendent d’une exposition maternelle. En conclusion, nous montrons que notre mélange de polluants possède une activité œstrogéno-mimétique. Ceci pourrait avoir une implication dans la compréhension du rôle des polluants environnementaux dans le développement des maladies métaboliques chez la femme en transition ménopausique / Menopause is a risk factor associated with the development of metabolic diseases. Because of estrogen deficiency, postmenopausal women may be at particular risk when exposed to chemical agents that can disrupt the endocrine system. To get more insight, we have developed a model of ovariectomized mice supplemented or not with estradiol (E2), and chronically exposed to a mixture of pollutants (TCDD, PCB153, DEHP and BPA) added to an obesogenic diet. Each pollutant was used at a dose close to the Tolerable Daily Intake dose, and the exposure had covered different stages of development, including or not the maternal period (gestation + lactation). Our results indicate that in diet-induced obesity, ovariectomy triggered weight gain, increase glucose intolerance and insulin resistance. This phenotype was associated with decreased estrogenic signaling in liver and deregulation of adipogenesis and inflammation in adipose tissues. Interestingly, the mixture of pollutants reduced the deleterious impact of ovariectomy on the metabolic phenotype, possibly through an alleviation of estrogenic signaling in both liver and adipose tissues. These data were not reproduced in mice not exposed to pollutants during the maternal life. In conclusion, the mixture of pollutants displayed some estrogeno-mimetic activity. This could have implications in understanding the role of environmental pollutants in the development of metabolic diseases in women in menopausal transition.

O processo inflamatório subclinico encontra-se associado à prevenção e controle de um agrupamento de fatores de risco (FR) nutricionais, entre eles, dislipidemia e aumento de depósito de gordura visceral. Indicadores nutricionais, inflamatórios e metabólicos parecem estar associados com o estilo de vida. A semente de linhaça tem sido reconhecida como um alimento rico em fibras e -3, entretanto, um novo constituinte de sua composição nutricional tem merecido atenção, pelo seu papel antiinflamatório e antioxidante. Este componente é chamado de lignanas, um polímero complexo e o principal constituinte não-carbohidrato de plantas vasculares. Está ligado a fibras de celulose, e é responsável por reforçar a estrutura das paredes celulares, o que previne o colapso das mesmas. Lignanas, em contato com a microflora intestinal humana transformam-se em enterolignanas, especialmente, enterodiol e enterolactona. O presente trabalho tem por hipótese que o teor de lignanas dietético pode interferir no perfil metabólico, e alterar fatores de riscos envolvidos no estado nutricional, e consequentemente na saúde. O conhecimento de que diferentes características na composição nutricional de macronutrientes da dieta poderiam modificar o perfil inflamatório, independentemente, da presença das enterolignanas provenientes da semente de linhaça também constituíram o objetivo deste estudo. Por 42 dias, foram avaliados 52 funcionários, pertencentes ao sexo masculino, com idade média de 37± 9 anos, de uma indústria de grande porte, na cidade de Itu-SP. Os voluntários foram divididos em 4 grupos de pesquisa, sendo, um grupo controle, e três grupos com dietas isocalóricas e diferentes proporções no % de carboidratos (CH), e acréscimo de semente de linhaça em pó ou arroz cru triturados (protocolo duplo cego). Foi preenchida ficha de coleta de informações sobre dados pessoais e conhecimento de fatores de risco (hipertensão, dislipidemia e diabetes), comportamentos de risco (tabagismo e sedentarismo) e antecedentes familiares. Foi também realizada avaliação clinico - laboratorial, no qual se obteve o registro de medidas antropométricas, medida da pressão arterial e coleta de sangue venoso em jejum de 12 h para avaliação de indicadores bioquímicos referentes à FR cardiovascular, tais como, colesterol total e frações (LDL-c e HDL-c), triglicérides, glicemia, insulina, Homa-beta e Homa-IR, ácido úrico, bem como, para avaliação de indicadores inflamatórios (Proteína C Reativa (PCR), Fator de Necrose Tumoral (TNF-) e Isoprostane Sérico), hormonais (Leptina e Adiponectina) e nutricionais (Enterodiol e Enterolactona séricas e urinárias). Observou-se que para redução significativa das medidas antropométricas estudadas e indicador de estresse oxidativo não houve diferenças entre os grupos que receberam intervenção dietética. Entretanto, para a melhora do perfil bioquímico, inflamatório, hormonal e nutricional, diferentes respostas foram encontradas. Os grupos que receberam dietas com redução de CH total (32% e 35%) mostraram benefícios, no que se refere ao perfil bioquímico, especialmente, colesterol total, LDL-c e ácido úrico, como também, para o perfil hormonal, referente aos níveis de adiponectina (p <0,05). Com relação aos níveis de PCR e TNF-, apenas os grupos que tiveram acréscimo de semente de linhaça na dieta apresentaram redução significativa (p<0,05). Para os níveis de triglicérides, somente o grupo com adição de semente de linhaça e 32 % de CH total apresentou diminuição significativa (p<0,05). Foi observado que com 32 % de CH total ingerido e adição de um alimento rico em lignanas constituiu-se uma estratégia nutricional relevante, para prevenção primária de fatores de risco metabólicos e controle da inflamação subclinica, o que pode contribuir na redução da morbi-mortalidade a eles associada. / The control of subclinical inflammatory process is associated with the prevention nutritional RF (risk factor), such as dislipidemia and the increase of visceral fat deposition. Nutritional, inflammatory and metabolic indicators seem also to related to life style. The linseed has been recognized as rich in fibers and -3. However, a new component in its nutritional composition has deserved the attention for its anti- inflammatory and antioxidant roles. This component is called lignans, a complex polymer and the main non-carbohydrate constituent of vascular plants. It is binded to cellulose fibers and is responsible for reinforcing cell walls structure, preventing them from collapsing. Plant lignans, in contact with the human intestinal flora, become enterolignans, specially enterodiol and enterolactone. This present work hypothetically that the amount of dietetic plant lignans interfere in the metabolic profile, altering the risk factors involved in the nutritional health state and consequently, the welfare state. Therefore, the objective of this study is to know if different diet nutritional composition characteristics can change the inflammatory profile, independently of the presence of enterolignans from the linseed. For 42 days, 52 male volunteers, average 37±9 years old, from a industrial city of Itu-SP, were evaluated. The volunteers were divided into 4 research groups; one control group and 3 groups on isocaloric diets with different proportion of carbohydrate (CH) and the addition of powdered linseed or ground raw rice (a double blind protocol). Personal data, RF (hypertension, dislipidemia and diabetes), habits (smoking and sedentary) and family antecedents were collected. A nutritional-laboratorial evaluation was performed in order to get anthropometric data; blood pressure checked and blood samples (after 12 hours fast) for total cholesterol and fractions (LDL-c and HDL-c ), triglycerides, glycemia, insulin, Homa-beta and Homa-IR, uric acid, inflammatory indicators (Reactive-C Protein (PCR) , Tumoral Necrosis Factor (TNF-a) and Seric Isoprostane ), hormonal (Leptin and Adiponectin), enterodiol and enterolactone seric and urinary enterolignins. It was observed a reduction of the studied anthropometric measures and for the oxidative stress indicators. A significant change occurred in the anthropometric measurements and the oxidative stress marker evaluated for all groups, but no difference among them was noted. On the other hand, biochemical, inflammatory, hormonal and nutritional profile significant differences among groups was observed. The groups that received diets with the reduction of the total CH (32% and 35% ) showed improvements in the biochemical profile, specially in the total cholesterol, LDL-c and uric acid, as well as the hormonal profile, in the levels of adiponectin (p< 0,05) . The levels of PCR and TNF-a, only the groups that had the linseed, showed a reduction (p< 0,05). For the triglycerides levels, only the group with the addition of linseed and 32% of total CH showed a decrease. It was observed with 32% of CH and the addition of food lignans constitute a nutritional relevant strategy for the primary prevention of metabolic risk factors and control of subclinical inflammation, contributing to the reduction of the associated morbi-mortality.

L’endothélium est considéré comme le plus grand organe endocrine, et est impliqué dans plusieurs processus physiologiques tels que l’inflammation et l’angiogenèse. Toute altération fonctionnelle/structurale peut induire de nombreuses maladies (des maladies cardiovasculaires jusqu’aux maladies métaboliques). L’identification de nouveaux facteurs moléculaires permettant de faire un lien entre les maladies cardiaques et les désordres métaboliques est de grande importance. Les prokinéticines sont des hormones peptidiques largement distribuées dans les tissus des mammifères. Elles agissent comme des facteurs de survie/mitogéniques dans plusieurs types cellulaires, notamment les cellules endothéliales, et sont impliquées dans la régulation des activités biologiques tels que l’angiogenèse et la physiopathologie cardiaque et rénale. Les prokinéticines agissent via deux récepteurs couplés aux protéines G: PKR1 et PKR2. Récemment il a été montré que l’activation de PKR1 induit l’angiogenèse tandis que l’activation de PKR2 induit la fenestration des cellules endothéliales, et que l’inactivation de PKR1 entraîne des anomalies cardiaques et rénales.Le but de cette étude est de comprendre à quel point la signalisation endothéliale de PKR1 peut affecter la fonction des organes adultes in vivo. Pour cela, nous avons généré une lignée de souris dans laquelle le PKR1 est spécifiquement inactivé dans les cellules endothéliales. Les souris mutantes présentent des dysfonctionnements endothéliaux avec des anomalies cardiaques, rénales et métaboliques. Les résultats montrent que PKR1 pourrait être considéré comme une cible pour le traitement des maladies cardiaques, rénales et métaboliques. / Endothelium is the largest living organ which has important role in many biological functions such as maintaining vascular smooth muscle tone, angiogenesis, cell proliferation, tissue growth and regulating lipid oxidation. Abnormalities in endothelium cause a large wide range of diseases from cardiovascular diseases to metabolic disorders. This emphasizes the important need for in-depth study of molecular and cellular mechanisms involved in endothelial dysfunction. Prokineticins are peptidic hormones widely distributed in peripheral and various endocrine mammalian tissues. Prokineticins are involved in many biological functions including angiogenesis, gastro-intestinal motility and regulation of heart and kidney physiopathology. These regulatory peptides act via two G protein couple receptor, PKR1 and PKR1. Recent studies show that PKR1 is involved in angiogenesis however PKR2 induces fenestration in coronary endothelial cells. However the role of PKR1 regulation in endothelial cells in vivo has not been studied yet.The aim of this study is to understand to what extent endothelial-prokineticin signaling affect adult organ functions in vivo. We had generated mutant mice lacking PKR1 in endothelium, by Cre-loxP technology, and studied how inactivation of PKR1 in endothelial cells affects different peripheral organ functions. These mice exhibit endothelial dysfunction as observed by abnormal endothelial cell proliferation and endothelial dependent relaxations. Moreover these mice exhibit cardiac, renal and metabolic disorders. Results indicate that PKR1 can be target for treatment of cardiovascular and metabolic disorders.

Dietas com alto teor de amido podem causar distúrbios metabólicos como acidose ruminal e abscessos hepáticos em bovinos. No entanto, o mais comum é observar desempenhos inconsistentes, particularmente em zebuínos. Aditivos como ionóforos e virginiamicina isoladamente foram bastante estudados e os resultados são consistentes em demonstrar melhora no desempenho e alteração da fermentação ruminal com redução de distúrbios metabólicos. A virginiamicina também apresenta melhora no desempenho em monogástricos. Estudos recentes sugerem que a virginiamicina apresenta efeito aditivo ao do ionóforo. O objetivo deste trabalho foi avaliar os efeitos da combinação de virginiamicina e ionóforo sobre o desempenho, características de carcaça, amido e pH fecais e incidência de abscessos hepáticos em bovinos Nelore alimentados com dietas de alto concentrado. 72 novilhos Nelore, castrados, com peso inicial de 278,9 kg, foram confinados em baias individuais por 61 dias após 15 dias de adaptação, recebendo dietas com dois níveis de concentrado (73 e 91%) e dois níveis de virginiamicina (0 e 15 mg/kg MS) em um esquema fatorial 2x2. Todas as dietas continham salinomicina (13 mg/kg MS). O consumo de matéria seca foi maior (P<0,01) para o tratamento com 91% de concentrado, tanto em kg/dia (8,96 vs. 7,79) como em porcentagem de peso vivo (2,07 vs. 1,83), assim como o consumo de energia metabolizável (26,20 vs. 19,86 Mcal/dia; P<0,01) e o ganho de peso (1,79 vs. 1,43 kg/dia; P<0,01). A eficiência alimentar apresentou tendência de aumento para o tratamento com mais concentrado (202,67 vs. 185,91 g/kg; P=0,08), no entanto, não houve diferença para eficiência de utilização de energia metabolizável (P=0,29), e teores de energia líquida para mantença (P=0,38) e ganho (P=0,38) das dietas. O consumo de matéria seca foi menor para o tratamento com virginiamicina, tanto em kg/dia (7,98 vs. 8,76; P<0,01) como em porcentagem de peso vivo (1,88 vs. 2,01; P=0,01), assim como o consumo de energia metabolizável (21,97 vs. 24,09 Mcal/dia; P<0,01). No entanto, o ganho de peso diário não diferiu entre os tratamentos (P=0,66). Os animais tratados com virginiamicina apresentaram maior eficiência alimentar (206,00 vs. 182,58 g/kg; P=0,02), maior eficiência de utilização da energia metabolizável (75,05 vs. 67,47 g/Mcal; P<0,05), maior teor de energia líquida para mantença (2,25 vs. 2,08 Mcal/kg MS; P=0,01) e para ganho (1,56 vs. 1,41 Mcal/kg MS; P=0,01). Animais tratados com mais concentrado apresentaram maior rendimento de carcaça (55,3 vs. 54,4%; P=0,02) e maior peso de gordura perirrenal e pélvica (8,06 vs. 7,15 kg; P=0,04). A inclusão de virginiamicina não afetou as características de carcaça. Não foram observados abscessos no fígado de nenhum dos animais. O teor de amido fecal foi maior (P<0,01) para o tratamento com 91% de concentrado (19,27 vs. 13,95%), mas não diferiu (P=0,40) em função dos níveis de virginiamicina. O uso combinado de ionóforo e virginiamicina melhorou a eficiência do uso de dietas de alto concentrado e alto amido para bovinos Nelore. / Zebu cattle fed high concentrate diets may present metabolic disorders, such as ruminal acidosis and liver abscesses. Also these animals display inconsistent performances in high starch diets. The use of feed additives such as ionophores and virginiamycin has been thoroughly investigated and the results are very consistent, with improvements in performance, changing ruminal fermentation and reducing the incidence of metabolic disorders. Virginiamycin has also consistent positive effects on performance in swine and poultry animals. Recent studies have suggested that virginiamycin and ionophore may have an additive effect. Thus, the objective of this research was to evaluate the effects of virginiamycin and ionophore in combination on performance, carcass traits, fecal starch, fecal pH and liver abscess incidence in Nellore cattle fed high concentrate diets. 72 castrated Nellore steers, with initial body weight of 278.9 kg, were fed in individual pens for 61 days after a 15 day adaptation period. Diets had two concentrate levels (73 and 91%) and two virginiamycin levels (0 and 15 mg/kg DM) in a 2x2 factorial. All diets had the ionophore salinomycin (13 mg/kg DM). Dry matter intake was higher (P<0.01) for the 91% concentrate treatment, in kg/day (8.96 vs. 7.79) and as percentage of body weight (2.07 vs. 1.83), as well as metabolizable energy intake (26.2 vs. 19.9 Mcal/day; P<0.01). Body weight gain (1.79 vs. 1.43 kg/day; P<0.01) was also higher. There was a trend for increased feed efficiency in the 91% concentrate treatment (202.7 vs. 185.9 g/kg; P=0.08), while no differences were observed for efficiency of metabolizable energy utilization (P=0.29) or dietary net energy content for maintenance (P=0.38) and gain (P=0.38). Dry matter intake was lower for the virginiamycin treatment, in kg/day (7.98 vs. 8.76; P<0.01) and as percentage of body weight (1.88 vs. 2.01; P=0.01), as well as metabolizable energy intake (22.0 vs. 24.1 Mcal/day; P <0.01). However, daily gain did not differ between treatments (P=0.66). Virginiamycin fed animals showed increased feed efficiency (206.0 vs. 182.6 g/kg; P=0.02), efficiency of metabolizable energy utilization (75.05 vs. 67.47 g/Mcal; P<0.05), and dietary net energy content for maintenance (2.25 vs. 2.08 Mcal/kg DM; P=0.01) and gain (1.56 vs. 1.41 Mcal/kg DM; P=0.01). Animals fed 91% concentrate diet had higher dressing percentages (55.25 vs. 54.42%; P=0.02) and heavier kidney and pelvic fat (8.06 vs. 7.15 kg; P=0.04). Virginiamycin levels did not affect carcass traits. Liver abscesses were not observed in these 3 year old animals. Fecal starch content was higher (P<0.01) for the 91% concentrate treatment (19.27 vs. 13.95%), but did not change (P=0.40) due to virginiamycin. These results suggest that the combined use of ionophore and virginiamycin is an efficient tool in high concentrate and high starch diets for Nellore cattle.

In Europa leiden etwa 120 Millionen aller 18- bis 80-jährigen an einer Hörbehinderung (16% Prävalenz). Die altersassoziierte Innenohrschwerhörigkeit (Age related hearing impairment, ARHI) hat damit Anschluss gefunden an die großen Volkskrankheiten: Ischämische Herzerkrankungen (5,9 %), Demenz (5,8 %), Diabetes mellitus und Cerebralen Erkrankungen (je 5,5 %). Der erste Teil der Arbeit befasst sich mit der Theorie des Hörens und seinen Störungen, der cochleären Durchblutung, den metabolischen Risikofaktoren, der Molekularbiologie und Epigenetik des Innenohres. Obwohl in der Literatur weitgehend Konsens über den Einfluss exogener Faktoren auf ARHI herrscht, wie Diabetes mellitus, Herz-Kreislauferkrankungen (Framingham Studie und CVD), Hypertonie, Rauchen, Body mass index (BMI) und genderspezifische hormonelle Einflüsse, gibt es kaum HNO-ärztliche Studien dazu. Der zweite Teil geht in einer Praxis-Fallstudie an 202 Patienten folgenden Fragen nach: 1. Gibt es zwischen metabolischen Erkrankungen, wie Adipositas, Störungen des Fett- und Glucosestoffwechsels, der Hypertonie und der Zunahme von Innenohrschwerhörigkeiten im Erwachsenenalter klinische Zusammenhänge? 2. Könnte eine erste Ohrsymptomatik Prädikator einer okkulten pathologischen Stoffwechselveränderung sein? 3. Gibt es Möglichkeiten einer Prävention? Im Ergebnis zeigten sich bei Patienten mit ARHI im Verhältnis Männer/Frauen: Störung des Glukosestoffwechsels bei 97 %/84 %, Lipidstoffwechselstörungen bei 76 %/85 %, eine Homocysteinämien bei 70 %/62 %, ein Anstieg des Plasminogen-Aktivator-Inhibitors (PAI) bei 65 %/38 % und eine Harnsäureerhöhung bei 48 %/15 %. Auch bei ersten Ohrsymptomen (Ohrdruck, Tinnitus, einzelne Frequenzsenken) fanden sich in jedem einzelnen Fall metabolische Veränderungen, die entweder allein oder in unterschiedlichen Kombinationen vorlagen. Männern mit metabolisch initiierten KHK-Erkrankung haben als Prädikator häufig eine erektile Dysfunktion. Erste auditorische Symptomen könnten Prädikator okkulter metabolischer Störungen bei Männern und Frauen sein. Anamnestisch unklare Innenohrprobleme sollten daher auf Stoffwechselstörungen untersucht werden, da diese Risikofaktoren darstellen, die leitlinienbasiert zu therapieren sind. Eine Prognose ist in keinem Einzelfall möglich, da die Folgen auf die molekularbiologischen Prozesse im Innenohres individuell nicht erkennbar sind. Doch sahen wir bei einigen Patienten deutliche Verbesserungen der Symptome und einen Hörschwellenanstieg. Andere Patienten zeigten innerhalb des Beobachtungszeitraums keine Änderung der audiogenen Symptomatik. Behandlungen über einen längeren Zeitraum an einer größeren Fallzahl wären daher sinnvoll. Die Ergebnisse könnten als erste sekundärpräventive Behandlung betrachtet werden, die im Einzelfall eine Hörgeräteversorgung entbehrlich machte. Sie stellen einen Beitrag zu Pfisters Empfehlung dar, mit den vorhandenen Erkenntnissen exogener Risikomechanismen, dringend neue therapeutische Präventionsstrategien für die HNO-ärztliche Tätigkeit zu entwickeln, die die neurale und molekulare Basis anstelle der Symptome der Erkrankung fokussieren.

L’acétylation des résidus de glucosamine terminaux par la N-acétyltransférase lysosomale (HGSNAT) est une étape essentielle de la dégradation catabolique de l’héparan sulfate. Des défauts dans cette réaction causent une maladie de surcharge lysosomale autosomale récessive rare : le désordre de Sanfilippo type C (SFC). À ce jour, 54 mutations ont été rapportées chez des patients SFC, incluant 13 mutations des sites d’épissage, 11 insertions et délétions, 8 mutations non-sens, 18 mutations faux-sens et 4 polymorphismes, avec différentes manifestations phénotypiques. Nous avons identifié 10 d’entre elles et effectué une étude exhaustive portant sur l’éventail des mutations SFC, leur distribution dans la population de patients, ainsi que leur impact potentiel sur la structure de la HGSNAT. Les erreurs d’épissage, les mutations non-sens, les insertions et les délétions devraient toutes entraîner un ARN non fonctionnel qui est rapidement dégradé par des mécanismes de contrôle qualité cellulaire. Les 4 polymorphismes identifiés sont des changements d'acides aminés qui ne modifient pas l'activité enzymatique, la glycosylation ou la localisation et n'ont donc pas de signification au niveau clinique. Au niveau des enzymes, les polymorphismes sont des changements d’acides aminés qui n’affectent pas la fonction, mais dans un contexte d’acides nucléiques ils peuvent être considérés comme des mutations faux-sens. Les dix-huit mutations faux-sens qui ont été exprimées ont produit des protéines inactives, en raison d'erreurs dans leur repliement. Ceci expliquerait donc la progression sévère de la maladie chez les personnes porteuses de ces mutations. Les protéines mutantes mal repliées sont anormalement glycosylées et conservées dans le réticulum endoplasmique. La thérapie par amélioration de l’activité enzymatique par des chaperonnes est une option thérapeutique potentielle, spécifiquement conçue pour exploiter l'activité enzymatique résiduelle de mutants mal repliés, afin d’éliminer les substrats stockés. Nous avons démontré que le traitement de plusieurs lignées de fibroblastes de patients SFC avec le chlorhydrate de glucosamine, un inhibiteur spécifique de la HGSNAT, a partiellement restauré l’activité de l'enzyme mutante, fournissant une preuve de l’utilité future de la thérapie par des chaperonnes dans le traitement de la maladie de SFC. / The acetylation of terminal glucosamine residues by lysosomal N-acetyltransferase (HGSNAT) is an essential part of the catabolic breakdown of heparan sulfate. Defects in this reaction result in the rare autosomal recessive lysosomal storage disorder Sanfilippo syndrome type C (SFC). To date 54 mutations in SFC patients have been reported including 13 splice-site mutations, 11 insertions and deletions, 8 nonsense, 18 missense and 4 polymorphisms, with different phenotypic manifestations. We have identified 10 of them and conducted a comprehensive review discussing the spectrum of Sanfilippo C mutations, their distribution within the patient population as well as how the mutations could potentially affect the structure of HGSNAT. Splicing errors, nonsense mutations, insertions and deletions were all predicted to result in non-functional RNA which is rapidly degraded by cellular quality control mechanisms. The 4 identified polymorphisms resulted in amino acid changes which did not affect the enzyme activity, glycosylation or targeting and were therefore not clinically significant. Polymorphisms, in the context of enzymes are amino acid changes not affecting function, but in the context of nucleic acids can still be considered as missense mutations. Eighteen missense mutations were expressed and shown be inactive due to errors in protein folding providing an explanation for the severe disease progression seen in individuals with these mutations. Misfolded mutants were abnormally glycosylated and retained in the endoplasmic reticulum. Enzyme enhancement/chaperone therapy is a potential treatment option specifically designed to exploit the residual enzyme activity of misfolded mutants in order to clear stored substrates. We demonstrated that treatment of several fibroblast lines of SFC patients with a specific inhibitor of HGSNAT; glucosamine-hydrochloride partially rescued mutant enzyme activity providing a proof of principle for the future use of chaperone therapeutics in the treatment of SFC.

Il sistema immunitario è costituito da una varietà di cellule, molecole e processi biologici che interagiscono per prevenire le invasioni microbiche, riconoscere le molecole estranee ed eliminare le fonti esistenti di lesioni cellulari, ripristinando le normali funzioni tissutali una volta risolto il problema. L'immunità innata è la prima linea di difesa contro le invasioni di agenti patogeni. Nelle vacche da latte, il suo funzionamento subisce gravi alterazioni durante il periodo di transizione (TP). In questa fase è stata segnalata una compromissione delle funzioni delle cellule polimorfonucleate (PMN) correlate alla produzione di metaboliti reattivi dell'ossigeno (ROM), all’attività della mieloperossidasi (MPO), alla chemiotassi e alla fagocitosi. I PMN bovini hanno un alterata espressione dei geni codificanti per tali funzioni tra -1 e 2 settimane dal parto, rispetto al livello rilevato 4 settimane dopo il parto per gli stessi geni. La causa esatta delle disfunzioni immunitarie che si verificano nel periparto non è mai stata chiaramente identificata. In esse possono contribuire diversi fattori, principalmente imputati alle alterazioni metaboliche tipiche del periparto (cambiamenti nell’assetto ormonale, limitazione della risposta immunitaria materna al fine di mantenere la gravidanza, alterazioni nel bilancio energetico e stato di stress ossidativo). Tuttavia, la durata e l’entità delle disfunzioni immunitarie può aumentare qualora subentri uno stato di squilibrio fisiologico (PI). In tali condizioni, le alterazioni metaboliche del periparto sfuggono al controllo dei meccanismi omeostatici e omeoretici, ed una infiammazione sistemica è la conseguenza frequente di questo squilibrio. Lo stato infiammatorio sistemico è scatenato da un aumento dei livelli di citochine proinfiammatorie (PIC), che è collegato ad un aumento della temperatura corporea al parto, e che tipicamente inficia le funzionalità epatiche, modificando le priorità anaboliche dell'organo in fase di inizio lattazione. A seguito di tale slittamento, il fegato produce più α-globuline, note come proteine positive di fase acuta (+APP), cioè aptoglobina, ceruloplasmina e siero amiloide alfa (SAA). Al contrario, riduce la sintesi di albumina, retinol binding protein (RBP), paraoxonasi (PON) e lipoproteine, note come proteine negative di fase acuta (-APP), e sequestra minerali, quali zinco e ferro, dal flusso ematico. L'infiammazione porta all'attivazione dei PMN, mentre la ridotta competenza immunitaria comunemente riportata in TP è stata associata ad un effetto opposto sui leucociti. Pertanto, questi dovrebbero essere considerati come due fenomeni distinti, ma lo stato di PI potrebbe essere considerato un denominatore comune, direttamente correlato al rischio di patologie in avvio di lattazione. Le strategie nutrizionali per ottimizzare l'immunità delle vacche da latte durante il TP dovrebbero quindi essere focalizzate sulla riduzione del grado di PI correlato al parto. Tra tali strategie nutrizionali, dovrebbe essere presa in considerazione la corretta gestione delle fonti energetiche per adattarle alle variazioni dei fabbisogni. Inoltre, il profilo degli acidi grassi delle fonti lipidiche può contribuire nel modificare le funzioni immunitarie. Infine, la somministrazione di prodotti supplementari con attività antiossidanti o antinfiammatorie, così come di specie donatrici di gruppi metilici, potrebbero essere strategie utili a favorire la funzionalità immunitaria delle bovine durante il TP. In una prospettiva più ampia, sebbene strategie nutrizionali e supplementi possano talora mitigare le alterazioni immunitarie, possiamo concludere che l'adozione di pratiche volte a minimizzare il PI durante il periodo di transizione sia la strategia più efficace per prevenire le disfunzioni. Al fine di chiarire il legame tra le alterazioni che si verificano nel periparto e le disfunzioni immunitarie delle bovine da latte sono stati condotti tre esperimenti. Bovine di razza frisona sono state alloggiate in poste individuali a stabulazione fissa e monitorate regolarmente per le condizioni corporee (BCS), il peso (BW), l'assunzione di alimenti (DMI), la produzione di latte (MY) e il tempo di ruminazione. Campioni di sangue sono stati raccolti regolarmente per valutare un ampio profilo ematochimico e per testare le funzioni dei globuli bianchi mediante stimolazioni ex-vivo. Inoltre, la diapedesi dei PMN è stata testata in vivo mediante test della carragenina e sono stati raccolti campioni di rumine a 30 giorni dal parto (DFC). Il primo esperimento era volto a chiarire le cause dei cambiamenti metabolici che si verificano al momento della messa in asciutta, ed il contributo del livello produttivo in tali alterazioni. Infatti, i profondi cambiamenti nell’alimentazione, gli adattamenti gastrointestinali, del metabolismo e dei parametri immunitari che si verificano nelle bovine alla messa in asciutta sono note scatenare il rilascio di cortisolo, indurre segnali di infiammazione sistemica ed alterare il bilancio redox. Produzioni di latte elevate al momento della messa in asciutta hanno un ruolo nell'aggravare tali condizioni. Nel nostro studio, un gruppo di 13 bovine è stato asciugato a 55 giorni dalla data prevista per il parto. Gli animali sono stati divisi in due gruppi in base alla produzione media dell'ultima settimana di lattazione, assumendo un cut-off di 15 kg * d-1: bassa (LM; 6 animali) e alta produzione (HM; 7 animali). I dati sono stati sottoposti ad ANOVA utilizzando un modello per misure ripetute, assumendo il livello produttivo al termine della lattazione, il tempo e la loro interazione come effetti fissi. L'aumento delle quantità di fibra nella razione di asciutta ha ridotto la DMI e aumentato il tempo di ruminazione. La migrazione dei leucociti nella ghiandola mammaria per contribuire alla fase di involuzione ha ridotto la loro abbondanza nel sangue e aumentato la loro attività. Tale attivazione dei leucociti nella mammella ha aumentato l'abbondanza di specie reattive dell’azoto nel plasma e innescato un'infiammazione sistemica in tutti gli animali (aumento delle +APP e riduzione delle -APP). Tale infiammazione ha compromesso le funzioni epatiche (aumento delle concentrazioni di gamma-glutamil transferasi -GGT- bilirubina e fosfatasi alcalina -ALP-). Sia la produzione di specie dell’azoto che lo stato infiammatorio sistemico hanno contribuito all'esaurimento degli antiossidanti circolanti (gruppi tiolici -SHp-, tocoferolo, β-carotene, potere antiossidante ferrico riducente -FRAP- e capacità antiossidante contro specie reattive dell'ossigeno -ORAC-). Gli animali con una produzione più elevata alla messa in asciutta hanno mostrato la peggiore condizione, probabilmente per i più profondi cambiamenti metabolici che hanno affrontato dopo l'interruzione delle mungiture, e per la fase involutiva verosimilmente più dispendiosa. Questo studio evidenzia la messa in asciutta come una fase critica per gestire la salute delle vacche da latte, e suggerisce un potenziale legame della messa in asciutta con le alterazioni delle funzioni immunitarie che si verificano nel periparto. Nel secondo esperimento si sono cercati di identificare i cambiamenti del sistema immunitario che precedono l'insorgenza della chetosi, al fine di chiarire il loro ruolo nella comparsa della malattia. Pertanto, 13 bovine sono state monitorate tra -48 e 35 DFC e suddivise in due gruppi sulla base dei loro livelli plasmatici di beta idrossibutirrato (BHB): inferiore (CTR, 7 animali) o superiore a 1,4 mMol / L (KET; 6 animali). I dati sono stati sottoposti ad ANOVA utilizzando un modello per misure ripetute, assumendo lo stato di salute, il tempo e la loro interazione come effetti fissi. Le vacche KET hanno avuto una maggiore attivazione del sistema immunitario prima del parto (maggiori concentrazioni plasmatiche di PIC, MPO e specie ossidanti e maggiori produzione di interferone gamma in risposta alla stimolazione con Mycobacterium avium) alterazioni della funzionalità epatica (più alta concentrazione sanguigna di GGT) e minori minerali plasmatici. Elevati livelli plasmatici di NEFA, BHB e glucosio nelle vacche KET suggeriscono uno stato di insulinoresistenza e una marcata mobilizzazione del grasso corporeo durante il periodo di asciutta. Tali andamenti dei parametri relativi al metabolismo energetico durante l’asciutta sono stati associati alla riduzione della DMI al momento del parto e al peggioramento del bilancio energetico negativo ad avvio lattazione. Ciò ha causato a sua volta una riduzione di MY e accresciuto ulteriormente la mobilizzazione dei grassi in avvio di lattazione. Compromissione della funzionalità epatica e attivazione dei leucociti durante il periodo di asciutta hanno determinato una marcata risposta infiammatoria di fase acuta nelle vacche KET dopo il parto (maggiori concentrazioni di +APP minori concentrazioni di RBP), ed ulteriormente compromesso la funzionalità epatica (maggiori concentrazioni di glutammato-ossalacetato transaminasi -AST-GOT- e bilirubina). I leucociti delle vacche KET hanno mostrato ridotte funzioni infiammatorie dopo stimolazione ex-vivo con lipopolisaccaridi batterici (minore produzione di PIC e maggiore produzione di lattato). Queste alterazioni potrebbero essere guidate dall'azione combinata dei metaboliti legati alla mobilizzazione dei lipidi e delle azioni antinfiammatorie volte a prevenire un'infiammazione eccessiva. Ciò suggerisce che le alterazioni dei parametri immunitari osservate prima del parto siano altamente correlate con la probabilità di sviluppare chetosi in avvio di lattazione. Nel terzo esperimento è stato somministrato un prodotto immunostimolante dalla comprovata efficacia nel migliorare le funzioni leucocitarie degli animali immunodepressi e nel ridurre l'incidenza delle malattie infettive delle bovine ad inizio lattazione. La sua modalità di azione non è mai stata chiarita, e un’indagine approfondita sul suo effetto metabolico potrebbe evidenziarne l’efficacia anche nei confronti dei disordini metabolici del periodo di transizione. Pertanto, un gruppo di10 bovine è stato monitorato da -62 a 42 DFC. Il gruppo trattato (TRT, 5 animali) ha ricevuto 32,5 g di Omnigen-AF® (Phibro Animal Health Corporation) due volte al giorno (65 g d-1), mentre il gruppo di controllo (CTR, 5 animali) non ha ricevuto alcun supplemento. I dati sono stati sottoposti ad ANOVA utilizzando un modello per misure ripetute, assumendo il trattamento, il tempo e la loro interazione come effetti fissi. La somministrazione dell’immunostimolante alla messa in asciutta non ha influenzato BW, BCS, MY, composizione del latte e del fluido ruminale e nemmeno modificato la concentrazione di neutrofili del sangue. Tuttavia, ha aumentato il tempo di ruminazione e migliorato il metabolismo energetico dopo il parto (concentrazioni di NEFA e BHB inferiori). Le bovine TRT avevano maggiori concentrazioni ematiche di linfociti e i loro leucociti avevano una maggiore efficienza nel rispondere alla stimolazione con lipopolisaccaridi batterici (produzione di lattato inferiore e minore consumo di glucosio). Nonostante questi effetti positivi sulle cellule immunitarie, l'immunostimolante non ha influenzato le concentrazioni di +APP dopo il parto. Inoltre, l’immunostimolante ha ridotto le concentrazioni di albumina, PON e antiossidanti dopo il parto, suggerendo la compromissione di alcune funzioni epatiche negli animali trattati. Tuttavia, la mancanza di qualsiasi effetto sui biomarcatori di funzionalità (bilirubina) e danno epatico (GGT, AST-GOT, ALP) smentisce una reale compromissione delle attività epatiche a seguito del trattamento. Gli effetti positivi nel favorire il recupero delle funzioni del rumine, riducendo la mobilizzazione dei grassi corporei dopo il parto, suggeriscono che l'immunostimolante sia una strategia efficace nella prevenzione dei disturbi metabolici del periodo di transizione. / Immune system is made of a variety of cells, molecules and biological processes that interacts to prevent microbial invasions, recognize foreign molecules and eliminate existing sources of cellular injuries to restore tissues to their normal functions once problem has been solved. Innate immunity is the primary defense line against pathogens invasions. Its functioning typically undergoes severe alterations during transition period (TP) of dairy cows. An impairment of polymorphonuclear cells (PMN) functions related to reactive oxygen metabolites (ROM) production, myeloperoxidase (MPO) activity, chemotaxis and phagocytosis has been reported in this phase. Bovine PMN have an altered abundance in mRNA transcripts encoding for such functions between -1 and 2 weeks from calving, in comparison to the level found at 4 weeks after calving for the same genes. The exact cause of immune dysfunctions occurring in peripartum has never been clearly identified. Reduced immune competence could arise from the interaction of different factors affected from the typical peripartum trends (i.e. changes in endocrine asset, limitations of maternal immune responses against the allogeneic conceptus, alterations in energy balance and oxidative stress status). Nevertheless, its duration could be modified when peripartal changes exceed the control of homeorhetic and homeostatic mechanisms, leading to the physiological imbalance (PI) condition. Such a condition could also trigger the inflammatory-like status. It consists in a prepartal raise of pro-inflammatory cytokines (PICs) levels, that is linked to a raise in body temperature at calving, and that typically affects liver metabolism, implying severe losses in hepatic functions and a shift of anabolic priority of the organ in early lactation. The liver produces more α-globulins, known as positive acute phase proteins (APP), i.e. haptoglobin, ceruloplasmin and serum amyloid alpha (SAA). Conversely, it reduces the synthesis of albumin, retinol binding protein, paraoxonase (PON) and lipoproteins, known as negative APP and sequesters minerals, as zinc and iron, from blood flow. Inflammation lead to the activation of PMN, while the reduced immune competence commonly reported in TP has been associated to an opposite effect on leukocytes. Thus, these should be considered as two distinct phenomena, but they could arise from a common cause with a different magnitude and duration. Nutritional strategies to optimize dairy cow’s immunity during TP should be focused on reducing the PI degree related to calving, as this condition could be referred as a common denominator between immune dysfunction and diseases causes. Among such nutritional strategies, the correct management of energy sources to fit with altered requirements should be considered. Furthermore, fatty acids profile of lipid sources administered could also modify immune functions. Finally, the administration of supplementary products exerting antioxidant or anti-inflammatory activities, as well as methyl donors species, could be beneficial for dairy cows immunity in TP. In a wider perspective, although feed additives and nutritional strategy could be effective in mitigate immune alterations, we can conclude that adoption of proper management practices aimed to avoid PI condition in peripartal period of dairy cows could be the most effective strategy to prevent dysfunctions. Three experiments have been designed to elucidate the linkage between sudden changes occurring in peripartum and immune alterations in dairy cows. Throughout such experiments Holstein dairy cows were housed in tied stalls and monitored regularly for body condition score (BCS), body weight (BW), dry matter intake (DMI), milk yield (MY) and rumination time. Blood samples were collected regularly to assess a wide hematochemical profile and to test white blood cell functions through ex-vivo challenges. Furthermore, PMN diapedesis has been tested in-vivo through a carrageenan-skin test and rumen samples were collected at 30 days from calving (DFC). The first experiment was aimed in investigate the main causes of metabolic changes occurring at dry-off and the contribution of MY in such alterations. In fact, dry-off is related to deep changes in feeding behavior, gastro intestinal adaptations, metabolism and immune parameters in high-yielding cow’s career. Indeed, the release of cortisol, signals of systemic inflammation and altered redox balance have been reported immediately after milking interruption, and high MY have a role in aggravating such conditions. In our study, a group of 13 Holstein dairy cows were dried off at 55 days from expected calving day, and regularly monitored from -7 to 34 days from dry-off (DFD). Animals were retrospectively divided in two groups according to their average MY in the last week of lactation, assuming a cut-off of 15 kg*d-1: low MY (6 cows) and high MY (7 cows). Data were submitted to ANOVA using a mixed model for repeated measures including MY at dry-off, time and their interaction as fixed effects. Increased fiber amounts of dry ration reduced DMI and increased rumination time. Leukocytes migration into mammary gland to contribute in the involution phase decreased their abundance in blood at dry-off, and their activity. Such activation of leukocytes at mammary site increased the abundance of nitrogen species in plasma and triggered a systemic inflammation in all the animals, as reflected from increased concentrations of positive and reduced concentrations of negative APPs. Such inflammation impaired liver functions, as suggested from the increased gamma-glutamyl transferase (GGT), bilirubin and alkaline phosphatase (ALP) concentrations. Both the production of nitrogen species and the systemic inflammatory status contributed in the depletion of antioxidant system in blood (thiol groups -SHp-, tocopherol, β-carotene, ferric reducing antioxidant power -FRAP- and oxygen reactive antioxidant capacity -ORAC-). Animals with higher MY at dry-off showed the worst condition, likely for the deeper metabolic changes they faced at milking interruption, and to the greater amount of mammary parenchyma to be reabsorbed. This study highlights the dry-off as a thorny point to manage dairy cows’ health and depose for a relationship between dry-off and immune alteration that typically occurs at calving. The second experiment was aimed in investigate changes occurring in the immune system prior to ketosis onset to elucidate their role in disease occurrence. Thus, a group of 13 Holstein dairy cows were monitored from -48 to 35 DFC and retrospectively divided into 2 groups basing on their plasma BHB levels: lower (CTR; 7 cows) or higher than 1.4 mMol/L (KET; 6 cows). Data were submitted to ANOVA using a mixed model for repeated measures including health status, time and their interaction as fixed effects. KET cows had a greater activation of the immune system prior to calving (higher plasma concentrations of PICs, myeloperoxidase and oxidant species, and greater interferon gamma responses to Mycobacterium avium) impaired liver functions (higher blood concentration of GGT) and lower plasma minerals. High plasma NEFA, BHB and glucose levels in KET cows suggest an insulin resistance status and a marked mobilization of body fat occurring during dry period. They were also associated to reduced DMI around calving and worse negative energy balance in early lactation. This caused in turn reduced MY and increased fat mobilization in early lactation. Impairment of liver function and activation of leukocytes during the dry period accentuated the acute phase response in KET cows after calving (greater concentrations of positive APPs and lower concentration of retinol binding protein), further impairing liver function (higher blood concentrations of glutamate-oxaloacetate transaminase -AST-GOT- and bilirubin). Leukocytes of KET cows had reduced inflammatory functions after an ex vivo stimulation assay (lower production of PICs and greater production of lactate). These alterations on WBC could be driven by the combined action of metabolites related to the mobilization of lipids and of anti-inflammatory actions aimed to prevent over exuberant inflammation. This suggests that prepartal trends of immune parameters be highly related with the likelihood of developing diseases in early lactation. The third experiment consisted in the administration of Omnigen-AF (OAF), an immune stimulant that is effective in increasing leukocytes functions in immunosuppressed animals and in reducing incidence of infectious diseases in early lactating dairy cows. Its mode of action has never been elucidated, and a wider perspective of its metabolic effect could highlight its effectiveness in facing metabolic disorders of transition period also. Thus, a group of 10 Holstein dairy cows were divided into 2 groups: treated group (TRT; 5 cows) received 32.5 g of Omnigen-AF® (Phibro Animal Health Corporation) twice a day (65 g d-1) as top-dress on the morning and afternoon feeds, while control group (CTR; 5 cows) did not receive any supplementation. From -62 to 42 DFC animals were monitored regularly. Data were submitted to ANOVA using a mixed model for repeated measures including treatment, time and their interaction as fixed effects. Administration of OAF at dry-off did not affect BW, BCS, milk yield, milk and rumen fluid composition, and neither affected blood neutrophils concentrations. Nevertheless, it increased rumination time and improved the energy metabolism after calving (lower NEFA and BHB concentrations). TRT cows had an increased lymphocytes abundance at blood level, and their leukocytes had greater efficiency in facing biological stressors during the peripartum (lower lactate production and lower glucose consumption after a challenge with bacterial lipopolysaccharides). Despite these positive effects on immune cells, OAF did not affect the positive APPs concentrations after calving. A reduced abundance of albumin, PON and antioxidants also occurred with OAF after calving, suggesting some impairment of hepatic functions to occur. Nevertheless, the lack of any effect on main biomarkers related to liver function (bilirubin) and liver damage (GGT, AST-GOT, ALP) dismisses a real impairment of liver activities to occur with OAF. Positive effects in favoring the recovery of rumen functions, reducing mobilization of body fats after calving suggest OAF to be an effective strategy in preventing metabolic disorders of transition period.

Si l’obésité a longtemps été considérée comme protectrice pour le squelette, sa prise en charge nutritionnelle s’accompagne d’une perte osseuse. Dans le but d’étudier les mécanismes liant le tissu adipeux et le tissu osseux, des rats ont été nourris avec un régime riche en graisse et en sucre afin d’induire une obésité et ses complications métaboliques. Puis, ces rats ont suivi un programme de prise en charge de l’obésité associant un rééquilibrage nutritionnel à de l’activité physique. Des investigations de la densité, de la micro architecture trabéculaire, de la qualité corticale, des propriétés biomécaniques et des paramètres cellulaires du tissu osseux ont été réalisées sur le squelette des rats. L’obésité induite a amélioré les paramètres densitométriques et corticaux des rats. La prise en charge de l’obésité a entraîné des effets contrastés. Le rééquilibrage de l’alimentation n’a pas altéré la densité ni les paramètres biomécaniques et corticaux du tissu osseux des rats en dépit d’altérations de la micro architecture et des perturbations l’activité cellulaire osseuse. L’exercice a augmenté la densité minérale osseuse du tibia des rats. Nos résultats montrent que l’inclusion de l’exercice dans un programme de prise en charge de l’obésité permet de potentialiser la perte de masse grasse et de maintenir l’intégrité du squelette. En étudiant l’impact d’un régime obésogène et de la prise en charge de l’obésité sur le tissu osseux chez le rat, ce travail de thèse apporte sa contribution dans la compréhension des mécanismes liant le tissu adipeux et le tissu osseux. / While obesity has long been considered protective of bone tissue, its nutritional management is linked to a concomitant bone loss. In order to access these mechanisms, male rats were given a high fat / high sucrose diet to induce obesity and its resulting metabolic disorders. Then, these rats participated in a well balanced nutritional program combined, or not, with physical activity in order to treat their obesity. Investigations on density, trabecular micro-architecture, cortical quality, biomechanical properties and cellular parameters of bone tissue were performed on rat’s skeleton. Obesity had improved the bone density and cortical parameters of rats. Obesity management had induced mixed effects. The well balanced diet did alter neither the bone density nor biomechanical and cortical parameters despite the presence of alterations in the trabecular micro architecture and some disturbances of bone cellular activity. Exercise by itself had increased the tibia bone mineral density. Our results show that including exercise in obesity management allows increasing the fat mass loss and maintaining the skeleton integrity. By studying the impact of an obesogenic diet and obesity treatment on bone tissue in rats, this study brings its contribution to the understanding of the mechanisms linking adipose tissue and bone tissue.

<p>Domestication has led to genetic changes that affect quantitative traits in farm animals. Both candidate gene analysis using association tests and genome scans based on linkage analysis have been performed to understand the molecular basis underlying quantitative genetic variation in horses, pigs and chickens. To test a possible association of polymorphisms in the <i>PRKAG3</i> gene, previously found to be associated with excess glycogen content in pig skeletal muscle, with quantitative traits in the horse, the major coding part of the equine <i>PRKAG3</i> sequence was identified. Bioinformatic characterization of the equine <i>PRKAG3</i> gene was conducted. A single nucleotide polymorphism (SNP) causing a missense mutation (Pro258Leu) was found. Screening this SNP showed that the Leu258 allele was more frequent in breeds with heavy muscularity. To assess previously reported associations between polymorphisms in the <i>MC4R</i> gene and obesity-related traits further, we conducted linkage analysis between the <i>MC4R</i> locus and fatness-related traits using a Wild BoarxLarge White intercross. No significant association between segregation at the <i>MC4R</i> locus and fatness was detected in this pedigree. A genome scan of quantitative trait loci (QTLs) has been performed in an intercross between chicken lines divergently selected for growth. Divergent parental lines have been established by selecting for high and low 56-day body weight for over 40 generations. The selection has led to approximately a 9-fold difference in 56-day body weight between lines and resulted in correlated responses for a number of traits including appetite, immune response, body composition and metabolic traits. Phenotypic data on growth and other correlated traits were collected from more than 800 F2 individuals. Genome scans using 145 markers on 26 linkage groups have identified QTLs affecting growth and correlated responses to selection for 56-day body weight. No major QTL explaining a large portion of phenotypic variation in growth was revealed in this study. </p>