Background: African-Americans are a vulnerable population group with disproportionately elevated rates of type 2 Diabetes Mellitus (DM). Resistant starch is a promising food ingredient that has the potential to reduce the risk factors involved in the development of type 2 DM. To date, there is a dearth of published research studies on the effect of resistant starch on African-Americans who are at increased risk for type 2 DM. Objective: The major objective of this study was to determine if daily consumption of approximately twelve grams of high-maize™ 260 resistant starch (RS) added to bread improved glucose homeostasis by monitoring changes in fasting plasma glucose, fructosamine, hemoglobin A1c, insulin, glucagon-like peptide-1, C-reactive protein, homeostasis model assessment insulin resistant (HOMA- IR) and beta-cell function (HOMA-Beta), serum acetate, propionate, and butyrate levels. Design: A fourteen-week, randomized, double-blind, within-subject crossover design feeding study was carried out in African-American males (n=8) and females (n=7) at increased risk for type 2 DM who resided in Southwest Virginia. All participants consumed bread containing added RS or control bread (no added RS) for six-weeks. RS and control bread feedings were separated by a two-week washout period. Results: Fasting Plasma Glucose (FPG) levels were significantly lower (P = 0.0179) after six-week control bread feedings compared to baseline. FPG levels were also significantly lower (P < 0.0001) after two-week washout period than at baseline. FPG levels were significantly higher (P < 0.0001) after six-week resistant starch bread feeding than at washout. FPG levels due to consumption of resistant starch versus control bread approached significance (P = 0.0574). Fructosamine levels were significantly lower (P = 0.0054) after control bread and resistant starch bread (P < 0.0012) consumption compared to baseline. No significant differences were found in fructosamine levels due to resistant bread intake versus control (P = 0.9692). Mean baseline HbA1c levels were 6.9% (n=15). This value was slightly lowered to 6.79% (n=14) at the end of the fourteen-week study, although statistical significance was not found. Mean ± standard errors for HbA1c values were 6.9% ± 0.18% and 6.9% ± 0.14% at baseline for the sequence groups, resistant starch first (n=7) and control treatment first (n=8) groups, respectively. Mean± standard error HbA1c values were 6.7%± 0.27% and 6.9% ± 0.27% at the conclusion of fourteen-week study for sequence groups, resistant starch first group (n=7) and control treatment first group, respectively. Baseline mean and standard errors C-reactive Protein (CRP) levels for male and female combined results were 0.62 ± 0.16 mg/dL (n=15). Mean CRP levels were 0.53 ± 0.12 mg/dL for resistant starch bread and 0.64 ± 0.21 mg/dL for control bread feeding periods. No significant differences were found for treatment, gender, or sequence effects for C-reactive protein levels during the fourteen-week study (P > 0.05). Mean HOMA-IR levels following six-week resistant starch and control bread consumption decreased to normal values (> 2.5), although no significant differences were found for treatment (P = 0.5923). Conclusions: Eighty-seven grams of Hi- maize™ 260 Resistant Starch added to baked loaves of bread consumed by a free-living African-American population at increased risk for type 2 diabetes did not consistently show significance in all clinical indicators and biochemical markers assessed. On the basis of the evidence in this study we do not have evidence that this amount of resistant starch in this population's diet will prevent the onset of diabetes. However, results are suggestive that higher levels of resistant starch in a more controlled experiment could reduce clinical risk factors for type 2 diabetes. / Ph. D.

The Atlantic cod (Gadus morhua L.) fishery is of great economic importance to the Faroese economy. There are two separately managed cod stocks around the Faroe Islands, the Faroe Plateau and the Faroe Bank cod. Both have experienced dramatic decreases in size and informed management decisions are vital for both stock viability and exploitation. The stocks are geographically isolated by an 800 m deep channel and water temperatures are on average 1 – 2 ºC higher on the Faroe Bank than on the Faroe Plateau. There are clear phenotypic differences between the stocks; in particular, the markedly higher growth rate for the Faroe Bank cod has caught public and scientific attention. There is continuing debate regarding the relative importance of genetics and environmental contributions to the contrasting phenotypes. Analyses of reproductive parameters (field data and experimental captive spawnings) as well as analyses of microsatellite and single nucleotide polymorphism (SNP) markers were undertaken to better resolve the issue. Field data as well as data from experimental captive spawnings provided evidence of reproductive differences between Faroe Plateau and Faroe Bank cod. Peak spawning occurred earlier on the Faroe Plateau than on the Faroe Bank and this difference in timing of spawning was maintained in captivity. In particular, differences in sizes of eggs (average diameters of 1.40 and 1.30 mm for Faroe Plateau and Faroe Bank cod eggs, respectively) and indirect evidence of greater volumes spawned by the Faroe Bank females suggested stock differences with respect to egg size – egg number trade-off. It was hypothesised that the strategy adopted by cod on the Faroe Bank, with a higher number of smaller eggs, evolved in response to a more hostile environment (bare seabed and higher exposure to predators) experienced by early life stages in this area. Experimental captive spawnings with Faroe Bank cod showed a large interfamily skew in survival rates of cod eggs and fry. Egg size was identified as a useful indicator of survival rates in the egg stage, but egg survival rates could not be used to predict viability in later developmental stages, thus highlighting the importance of employing some sort of genetic monitoring of cod fry to ensure sufficient family representation in the progeny. While no tank effect was evident concerning fry survival, a significant tank effect was identified concerning body sizes of fry. Microsatellite data were analysed using large sample sizes of Faroe Plateau and Faroe Bank cod with the Faroe Plateau divided into two locations, Faroe Plateau North-East and Faroe Plateau West (cod from each of the two were known to belong to separate spawning grounds). Two Norwegian coastal cod samples were included as outlier populations. While no genetic differentiation was detected between the two Faroe Plateau locations, these analyses revealed a detectable, albeit relatively modest, degree of genetic differentiation between cod from the Faroe Plateau and the Faroe Bank (FST = 0.0014 and 0.0018; DJost_EST = 0.0027 and 0.0048; P < 0.0001 and P < 0.001 for the Faroe Plateau North-East – Faroe Bank and the Faroe Plateau West – Faroe Bank comparisons). These values were several times smaller than those between Faroese and Norwegian coastal cod (pairwise FST and DJost_EST values in the range of 0.0061 – 0.0137 and 0.0158 – 0.0386, respectively). Despite recent reductions in census population sizes for Faroe Plateau and, particularly, Faroe Bank cod, genetic diversity estimates were comparable to the ones observed for Norwegian coastal cod and there was no evidence of significant genetic bottlenecks. Lastly, data for one of the markers (Gmo132) indicated genotype-dependent vertical distribution of cod (as investigated for Faroe Plateau North-East cod). Contrary to some previously published studies, analysis of SNPs of two candidate genes for adaptive divergence, the hemoglobin gene Hb-ß1 and the transferrin gene Tf1, failed to detect differentiation between samples of Faroe Plateau and Faroe Bank cod analysed in this thesis. Of 3533 novel SNPs simultaneously discovered and genotyped by restriction-site associated DNA (RAD) sequencing, 58 showed evidence of genetic differentiation between Faroe Plateau North-East and Faroe Bank cod (P < 0.05). No single locus was fixed for different alleles between Faroe Plateau and Faroe Bank cod. A set of eight informative SNPs (FST values between Faroe Plateau and Faroe Bank samples > 0.25; P < 0.0005) were selected for validation in larger samples, that included cod from both Faroe Plateau areas and the Faroe Bank as well as Norwegian coastal and White Sea cod. Six out of the eight loci amplified successfully with a PCR-based method and there was 100 % concordance between genotypes of individuals screened by both techniques. Due to ascertainment bias, the SNPs should only be applied with caution in a broader geographical context. Nonetheless, these SNPs did confirm the genetic substructure suggested for Faroese cod by microsatellite analyses. While no genetic differentiation was evident between the two Faroe Plateau locations, significant genetic differentiation was evident between Faroe Plateau and Faroe Bank cod at five of the SNPs (FST values in the range of 0.0383 – 0.1914). This panel of five SNPs could confidently be used to trace groups of Faroe Plateau and Faroe Bank cod to their population of origin. In conclusion, multiple lines of evidence demonstrate that Faroe Plateau and Faroe Bank cod are truly two genetically distinct populations. While the findings contribute to a broader understanding of the biology and the genetics of Faroe Plateau and Faroe Bank cod, the novel SNPs developed may provide a valuable resource for potential future demands of i.e. genetic stock identification methods.

Ce mémoire est séparé en deux volets, tous deux axés sur la spectroscopie proche infra-rouge (NIRS) pour la quantification des paramètres hémodynamiques. La NIRS est principalement basée sur la mesure des coefficients d'absorption (μa) et de dispersion (μs’) des tissus afin de retrouver les concentrations d'oxy- et de déoxyhémoglobine dans le sang. L'imagerie à l'aide de la NIRS est basée sur le parcours des photons à travers le tissu biologique à différentes longueurs d'onde du spectre proche infra-rouge. Le premier appareil de NIRS dont il sera question est un appareil de spectroscopie résolue dans le temps. Ce type de système retrouve des concentrations absolues d'hémoglobine à l'aide d'un bandeau placé sur la peau d’un sujet, dans ce cas-ci sur le front. Le modèle d’analyse des données permet la séparation des contributions extra-cérébrales et cérébrales aux données. Cette méthode fournit des données plus exactes sur la saturation en oxygène du cerveau, par rapport à un modèle homogène où le signal est contaminé par les couches superficielles. Une étude sur les changements hémodynamiques cérébraux de jeunes adultes pendant une activité physique a été réalisée, et l’article en détaillant les résultats est transcrit au chapitre 2. Le chapitre 3 comprend un retour sur cette étude et aborde les possibilités de travaux futurs. La seconde partie de mes travaux s’est déroulée sous forme de stage en entreprise durant l’été 2016. Sous la supervision de Dennis Hueber, Ph. D., et Beniamino Barbieri, Ph. D., j’ai effectué des recherches portant sur un appareil de NIRS manufacturé par la compagnie ISS Inc. et dont un prototype se trouve actuellement dans le laboratoire de mon superviseur Mathieu Dehaes, Ph. D. Cet appareil combine deux modalités d’imagerie optique, soit la NIRS dans le domaine des fréquences et la spectroscopie de corrélation diffuse. Le chapitre 4 détaille les tâches que j’ai réalisées durant ce temps, de même que les résultats des analyses que j’ai effectuées. / This master’s thesis is separated in two phases, both focused on near infrared spectroscopy for the quantification of hemodynamic parameters. NIRS is based on the measure of absorption (μa) and scattering (μs’) coefficients of tissues in order to recover the oxy- and deoxyhemoglobin concentrations in the blood. Its results are based on the photon propagation in tissue at different near-infrared wavelengths. The first NIRS system used during my studies is a time-resolved spectroscopy system. This device allowed us to retrieve absolute hemoglobin concentrations using a headband placed over the subject’s skin and centered on their forehead. The data analysis model which we used allowed us to separate extra-cerebral and cerebral contributions of the signal. This method yielded quantitative absolute measures of cerebral oxygen saturation as opposed to the traditional homogenous model where the signal is contaminated by superficial layers. A study on cerebral hemodynamic changes in young adults during exercise was conducted, and the published article detailing its results is transcribed in Chapter 2. Chapter 3 includes a review of this study and discusses potential future works. The second part of my research consisted in an industrial internship during the summer of 2016. Under the supervision of Dennis Hueber, Ph. D., and Beniamino Barbieri, Ph. D., I have worked on a NIRS device manufactured by ISS Inc., a prototype of which is currently in the laboratory of my supervisor Mathieu Dehaes, Ph. D. This device combines two NIRS modalities: frequency-domain NIRS and diffuse correlation spectroscopy. Chapter 4 details the work I have performed at ISS and the results of my research and analysis.

Previous studies that utilized sodium lactate infusion did not use resistance exercise protocol or analyzed muscle biopsies, or performed sex specific analysis. Aim: We initiated a project where resistance exercise was performed with low and high levels of lactate, acquired by venous lactate infusion where the specific aim of this study was to investigate and chart the physiological characteristics of sodium lactate infusion during a bout of resistance exercise on whole group level and sexes separated Method: A randomized, placebo controlled, cross-over design was implemented where male (n = 8) and female (n = 8) subjects accustomed to resistance exercise visited the laboratory three times for preliminary testing and training familiarization. In the following two experimental trials subjects arrived in an overnight fasted state. A resting state muscle biopsy was extracted from m. vastus lateralis and repeated blood samples were initiated which followed by 20 minute of baseline infusion of either infusate in resting state at 0.05 mmol/kg/min infusion rate with additional bolus doses during subsequent exercise. Following a brief warm up, unilateral knee-extensions (6 x 8-10 reps at 75% of 1-RM) were performered with or without venous infusion of sodium lactate, with volume matched saline as control. Exercise load and volume were matched between trials. Four additional biopsies were extracted at post-exercise, recovery period, and 24-hour post-exercise. Results: Sodium lactate infusion vs saline infusion respectively during resistance exercise yielded significantly higher blood lactate with sodium lactate (6.78 ± 0.33 mmol/l vs 2.99 ± 0.17 mmol/l), plasma lactate (8.86 ± 0.39 mmol/l vs 4.39 ± 0.22 mmol/l), blood sodium (143 ± 0.4 mmol/l vs 142 ± 0.3 mmol/l), blood pH (7.42 ± 0.01 vs 7.34 ± 0.01), but lower blood potassium (3.9 ± 0.1 mmol/l vs 4.2 ±  0.1 mmol/l), all  immediately following exercise. Sodium lactate infusion elicited main effect of trials and muscle lactate increased from baseline (8.5 ± 0.9 mmol·kg-1 dw vs 7.0 ± 0.6 mmol·kg-1 dw) to post-exercise (31.5 ± 2.8 mmol·kg-1 dw vs 26.9 ± 3.2 mmol·kg-1 dw) with sodium lactate and saline infusion respectively. Blood glucose, hemoglobin and muscle pH was not affected by sodium lactate infusion. Conclusions: Utilization of the sodium lactate infusion method during a bout of resistance exercise may be used as tool to effectively increase blood/plasma lactate and, to lesser extent, muscle content of lactate. However, a concomitant slightly alkalizing effect of blood likely will occur. / Tidigare studier som använt natriumlaktat infusion använde inte styrketräningsprotokoll, eller analyserade muskelbiopsier eller utförde könsspecifika analyser. Syfte och frågeställningar: Vi initierade ett projekt där styrketräning utfördes med låga eller höga nivåer av laktat som erhölls genom venös natriumlaktat infusion med det specifika syftet att undersöka och kartlägga fysiologisk karakteristiska av naturiumlaktat infusion under styrketräningsövning på helgrupps- och könsseparerad nivå. Följande frågeställningar inrättades; hur påverkar natriumlaktat infusion under styrketräning helblod- och plasma laktat, glukos, natrium, kalium, plasma volym genom hemoglobin och hematokrit, blod pH, muskellaktat- och muskel pH samt om skillnader i respons finns efter att könsspecifika analyser utförts på dessa variabler. Metod: En randomiserad, placebokontrollerad cross-over design implementerades där styrketräningsvana män (n = 8) och kvinnor (n = 8) besökte laboratoriet tre gånger för preliminäraför tester och träningsfamiliarisering. I efterföljande två experimentella försök anlände försökspersonerna i ett över nattligt fastande tillstånd. En baslinje biopsi extraherades från m. vastus lateralis och repeterade blodprover initierades med efterföljande 20 minuter av baslinje infusion av endera infusat i vilotillstånd med 0.05 mmol/kg/min infusionshastighet med ytterligare bolusdoser under efterföljande träning. Efter en kort uppvärmning utfördes unilaterala knäextensioner (6 x 8-10 reps vid 75% av 1-RM) med eller utan venös infusion av natrium laktat, med volymmatchande saltlösning som kontroll. Träningsbelastning och volym matchades mellan försök. Ytterligare fyra biopsier extraherades vid efter-träning, återhämtningsperiod, och efter 24 timmar. Resultat: Natriumlaktat respektive saltlösnings infusion under styrketräning gav signifikant högre blodlaktat med natriumlaktat infusion (6.78 ± 0.33 mmol/l mot 2.99 ± 0.17 mmol/l), plasmalaktat (8.86 ± 0.39 mmol/l mot 4.39 ± 0.22 mmol/l), blodnatrium (143 ± 0.4 mmol/l mot 142 ± 0.3 mmol/l), blod pH (7.42 ± 0.01 mot 7.34 ± 0.01), men lägre blod kalium (3.9 ± 0.1 mmol/l mot 4.2 ± 0.1 mmol/l), alla direkt efter träning. Natriumlaktat infusion framkallade huvudeffekt av försök och muskellaktat ökade från baslinje (8.5 ± 0.9 mmol·kg-1 dw mot 7.0 ± 0.6 mmol·kg-1 dw) till efter-träning (31.5 ± 2.8 mmol·kg-1 dw mot 26.9 ± 3.2 mmol·kg-1 dw) med natriumlaktat respektive saltlösnings infusion. Blodglukos, hemoglobin och muskel pH påverkades inte av natriumlaktat infusion. Slutsats: Användande av natriumlaktat infusion som metod under styrketräning kan effektivt användas som verktyg för att höja blod/plasma laktat, och i mindre utsträckning, muskellaktat. Emellertid är samtidig alkalisering av blod en sannolik följd. / Potential sex differences in the molecular response to resistance exercise with lactate infusion

La Tomographie Optique Diffuse (TOD) est désormais une modalité d’imagerie médicale fonctionnelle reconnue. L’une des applications les plus répandues de cette technique est celle de l’imagerie fonctionnelle cérébrale chez l’Homme. En effet, cette technique présente de nombreux avantages, notamment grâce à la richesse des contrastes optiques accessibles. Néanmoins, certains verrous subsistent et freinent le développement de son utilisation, spécialement pour des applications chez l’Homme adulte en clinique ou dans des conditions particulières comme lors du suivi de l’activité sportive. En effet, le signal optique mesuré contient des informations venant de différentes profondeurs de la tête, et donc de différents types de tissus comme la peau ou le cerveau. Or, la réponse d’intérêt étant celle du cerveau, la réponse de la peau peut dégrader l’information recherchée. Dans ce contexte, ces travaux portent sur le développement d’un nouvel instrument de TOD permettant d’acquérir les dimensions spatiale, spectrale et de temps de vol du photon de façon simultanée, et ce à haute fréquence d’acquisition. Au cours de cette thèse, l’instrument a été développé et caractérisé sur fantôme optique. Ensuite, il a été validé in-vivo chez l’Homme adulte, notamment en détectant l’activité du cortex préfrontal en réponse à une tâche de calcul simple. Les informations multidimensionnelles acquises par notre système ont permis d’améliorer la séparation des contributions des différents tissus (Peau/Cerveau). Elles ont également permis de différencier la signature de la réponse physiologique de ces tissus, notamment en permettant de détecter les variations de concentration en Cytochrome-c-oxydase. Parallèlement à ce développement instrumental, des simulations Monte-Carlo de la propagation de la lumière dans un modèle anatomique de tête ont été effectuées. Ces simulations ont permis de mieux comprendre la propagation de la lumière dans les tissus en fonction de la longueur d’onde et de valider la pertinence de cette approche multidimensionnelle. Les perspectives de ces travaux de thèse se dirigent vers l’utilisation de cet instrument pour le suivi de la réponse du cerveau chez l’Homme adulte lors de différentes sollicitations comme des stimulations de TDCS, ou en réponse à une activité sportive. / The Diffuse Optical Tomography (DOT) is now a relevant tool for the functional medical imaging. One of the most widespread application of this technic is the imaging of the human brain function. Indeed, this technic has numerous advantages, especially the richness of the optical contrast accessible. Nevertheless, some drawbacks are curbing the use of the technic, especially for applications on adults in clinics or in particular environment like in the monitoring of sports activity. Indeed, the measured signal contains information coming from different depths of the head, so it contains different tissues types like skin and brain. Yet, the response of interest is the one of the brain, and the one of the skin is blurring it. In this context, this work is about the development of a new instrument of DOT capable of acquiring spatial and spectral information, as well as the arrival time of photons simultaneously and at a high acquisition speed. During the PhD thesis the instrument has been developed and characterised on optical phantoms. Then, it has been validated in-vivo on adults, especially by detecting the cortical activation of the prefrontal cortex, in response to a simple calculation task. Multidimensional information acquired by our system allowed us to better distinguish between superficial and deep layers. It also allowed us to distinguish between the physiological signature of those tissues, and especially to detect the variations of concentration in Cytochrom-c-oxydase. Concurrently to this experimental work, Monte-Carlo simulation of light propagation in a model off a human head has been done. Those simulations allowed us to better understand the light propagation in tissues as function as their wavelength, and to validate the relevance of our multidimensional approach. Perspectives of this work is to use the developed instrument to monitor the brain’s response of the Human adult to several solicitations like tDCS stimulation, or sports activity.

L’anémie est fréquente chez les patients pédiatriques en postopératoire de chirurgie cardiaque. Malgré le nombre important de patients transfusés, le taux d’hémoglobine (Hb) pour lequel les bénéfices surpassent les risques est inconnu chez ces patients. Récemment, Lacroix et al. ont démontré qu’une stratégie transfusionnelle restrictive n’était pas inférieure à une stratégie libérale en ce qui concerne le développement ou la progression du syndrome de défaillance multiviscérale (SDMV) et la mortalité chez les patients de soins intensifs pédiatriques (SIP).Devant le manque d’évidence, une analyse de sous-groupes des patients en postopératoire de chirurgie cardiaque de l’étude Transfusion Requirements in Pediatric Intensive Care (TRIPICU) a été réalisée. L’objectif de cette étude était de déterminer l’impact d’une stratégie transfusionnelle restrictive comparée à une stratégie libérale sur l’acquisition ou l’aggravation du syndrome de défaillance multiviscérale (SDMV) chez les enfants en postopératoire de chirurgie cardiaque. Cette étude n’a pas démontré de différences statistiquement, ni cliniquement significatives du nombre de patients ayant acquis ou aggravés un SDMV, ni des issues secondaires entre les stratégies transfusionnelles restrictive et libérale. L’analyse de sous-groupes permet de générer une hypothèse de recherche et les résultats devraient être confirmés par un essai randomisé contrôlé. / Anemia is frequent in pediatric patients following cardiac surgery. Despite frequent transfusions, the optimal hemoglobin threshold where benefits surpass risks is still unknown for these patients. Recently, Lacroix et al. showed that a restrictive transfusion strategy was not inferior to a liberal strategy concerning the development or progression of multiple organ dysfunction syndrome (MODS) and mortality in pediatric intensive care patients. In the absence of evidence, the aim of this study was to determine the impact of a restrictive versus a liberal transfusion strategy on new or progressive multiple organ dysfunction syndrome (MODS) in children following cardiac surgery. We conducted a subgroup analysis of the postoperative cardiac surgery patients of the Transfusion Requirements in Pediatric Intensive Care Unit (TRIPICU) study. Our study showed no statistically and clinically significant differences in the number of patients who acquired or worsened MODS, nor secondary outcomes between a restrictive and a liberal transfusion strategy. This subgroup analysis generates a research hypothesis that should be confirmed by a randomized controlled trial.

Fundação de Amparo a Pesquisa do Estado de Minas Gerais / Doutor em Genética e Bioquímica / CHAPTER II: Purpose: We performed a meta-analysis of randomized trials to assess the effect of metformin on inflammatory markers and metabolic parameters in subjects with diabetes. Methods: We performed comprehensive searches on NCBI, Cochrane, Science Direct databases from 1966 to Jun of 2015. We included randomized trials of at least 4 weeks duration that compared groups with diabetes before and after the treatment with metformin or metformin plus other drugs, and evaluated body mass index, blood glucose, HbA1c and inflammatory parameters such as C-reactive protein, tumor necrosis factor and adiponectin. Results: Pooled results of the 26 trials, with 1760 participants at the end of treatment reduce BMI in 0.9% p=0,0043, as well as, decrease of blood glucose level [SMD -0,411 mg/dL, 95%CI -0,463 to -0,369, I2= 56.62%], HbA1c [SMD -0.479%, 95%CI -0,568 to -0,390, I2= 55.02%], CRP levels [SMD -0,274mg/dL, 95%CI -0,419 to -0,129, I2= 72.78%], TNFα concentration [SMD -0,103pg/ml, 95%CI -0,514 to 0,309, I2= 87.67%] and increase of adiponectin [SMD 0,171μg/ml, 95%CI 0,098 to 0,440, I2= 81.09%] compared with pretreatment. Conclusion: The long-treatment with metformin monotherapy or metformin plus other drugs improves metabolic parameters and induced changes in inflammatory markers in diabetic subject. CHAPTER III: Background: Metformin increases insulin sensitivity by decreasing hepatic glucose production and increasing glucose disposal in skeletal muscle. However, modulation of inflammatory response and CaMKKβ/AMPK/Myosin V activation in gastrocnemius muscle by metformin treatment has not been demonstrated in hypoinsulinemic diabetic rats. Objective: The present study investigated how the metformin improve insulin sensitivity in skeletal muscle of hypoinsulinemic diabetic rats. Methods: Diabetes was induced by streptozotocin (45 mg/kg, intraperitoneally) 10 days prior treatments. On 11th day, diabetic rats were treated with metformin (500 mg/kg, oral gavage), insulin (2U at 08:00 h and 4U at 17:00 h, subcutaneously) or untreated. After 20 days, glycemia was measured and insulin sensitivity was determined by KITT. Serum Insulin, GLUT4, IRSthr, inflammatory markers (NF-κB, IκB, TNF-α and p-JNK) and CAMKK, AMPK and Myosin V in gastrocnemius muscle were determined by ELISA. Results: As expected, insulin and metformin improved the insulin sensitivity. Besides, metformin treatment promoted reduction in inflammatory response mediated by NF-κB, IκB, TNF-α and p-JNK, and that was accompanied by increased CaMKKβ/AMPK/Myosin V/GLUT4 pathway activity in gastrocnemius muscle of diabetic rats. Conclusion: Our findings suggest that metformin induces significant reductions in several inflammatory markers in skeletal muscle of diabetic rats. Metformin-induced increase in CaMKKβ/AMPK/Myosin V/GLUT4 pathway activity was associated with higher insulin sensitivity. CHAPTER IV: Diabetes is characterized by a proinflammatory state which can activate TLR2 and TLR4, and these receptors could induce NF-κB and JNK activation in skeletal muscle. In this study, we investigated the inflammatory and apoptotic signaling pathways triggered by TLRs/NF-κB and JNK activation in skeletal muscle of diabetic rats treated with metformin before and after an insulin tolerance test. Metformin treatment decreased p-JNK and NF-κB, and increased IκB concentrations. This attenuation leads to a decrease of TNFα and CXCL1/KC, and an increase of p-AMPK, BAX and Bcl2 concentration. Furthermore, KITT revealed an improvement of the insulin sensitivity in the diabetic rats treated with metformin. In addition, metformin was not capable of attenuating the changes in the inflammatory pathway triggered by insulin injection as the increase of TNFα and TLR4 in metformin treated rats, and IκB, CXCL1/KC, TNFα and p-AMPK increase in the untreated group. Taken together, these results point out that metformin may attenuate the activation of the inflammatory pathway TLRs/NF-κB/TNFα/CXCL1/KC and the apoptotic signaling BAX/Bcl2/p-JNK, which could be accompanied by a reduction of the inflammatory damage caused by hyperglycemia and an improvement of insulin sensitivity in diabetic rats.

Sheep breeding is nowadays a developing branch of agriculture again. There is a lot of different breed and they react distinctly on the same conditions of the enviroment. Sheep of breeds charollais, suffolk, šumavská ovce and valaška bred in similar conditions were observed in spring and autumn of years 2007 and 2008. In these seasons were taking blood samples (from {$\pm$}7{--}24) ewes and lambs and were analyzed in hematology laboratory. There were determined haemoglobin level, haematocrit indicator, erytrocytes and leucocytes levels, glucose, cholesterol, triglycerides levels, urea and plasmatic proteins, activity of ALP and GMT enzymes, phosphor, calcium, magnesium, zinc and copper levels. The main objective of this project was determine seasonal changes in observed parametres. Next objectives were determine different changes in blood parametres in different breeds and different aimes of breeds. There were recognized that all the observed breeds don`t react the same way on similar conditions. There were not provably determined seasonal changes in observed parametres, but average Hb level was higher in autumn than in spring. Urea level was conversely higher in spring season than in autumn. The demostrable fact is, that the similar conditions induce different answers not only in different breeds but also in different aimes of breeds.

To address the challenge of accurate, low cost and robust biosensors for diabetes management and early detection of diabetes complications, we have developed novel, robust sensing chemistry (or receptors) for electrochemical POC biosensors. The biosensors have been developed for the bio-markers associated with diabetes management such as glycated haemoglobin (HbA1c), glycated albumin, glucose, biomarkers associated with diabetes complications such as microalbuminuria, urine creatinine and albumin-to-creatinine ratio (ACR) and biomarkers associated with anaemia and malnutrition conditions such as haemoglobin and serum albumin. For haemoglobin detection, a new POC bio sensing technique has been developed based on Aza-heterocyclic chemicals. The repeatability and accuracy of the biosensor have been tested on real pathology samples. The glycated form of haemoglobin, called glycated haemoglobin or HbA1c, is the gold standard test in diabetes management as it gives the 90-days average blood glucose value. We demonstrate a simple method for electrochemical detection of HbA1c by combining bosonic affinity principle along with aza-heterocyclic receptors. The technique has been verified on the real clinical patient samples. Albumin is the most abundant protein in the human blood. Human serum albumin (HSA) is either alone or an associative biomarker in several chronic diseases like necrosis, nephrosis, hepatitis, malnutrition, arthritis, immune disorders, cancer, diabetes and in some severe infections. In pathology laboratories, the serum albumin is usually tested on serum samples and not in whole blood samples. Since albumin is not a metalloproteinase, it is very difficult to develop electrochemical POC biosensor. We have developed a novel technique for the electrochemical detection of serum albumin in whole blood samples, by exploiting its binding property with redox active copper salts. The accuracy of technique has been verified on both real human blood plasma as well as whole blood samples. Glycated albumin, which is the glycated form of serum albumin, is emerging as a novel biomarker for diabetes management, as it gives the average blood glucose value of 15-20 days. It is also extremely useful in chronic kidney disease patients and patients with hemoglobinopathies where HbA1c can give the erroneous results. By combining the copper chemistry along with bosonic affinity principle, we present the first ever demonstration of glycated albumin sensing. Instant blood glucose monitoring is an integral part of diabetes management. Most of the glucometers available in the market are based on glucose oxidase enzyme. We have demonstrated a low cost non-enzymatic electrochemical technique for blood glucose detection using alkaline methylene blue chemistry. The accuracy of the technique has been verified on real human blood plasma samples. Glucometer is one of the most easily available POC biosensor and a useful tool for diabetes population. India has second largest diabetes population in the world. To analyse the accuracy of the POC glucometers which are available in Indian market, a comprehensive study was conducted. The results were compared with clinical accuracy guidelines using exhaustive statistical analysis techniques. The shortcomings of the commercial glucometers are elucidated, regarding different international standards. Diabetic nephropathy is one of the major diabetes complications and is the primary cause of chronic kidney disease (CKD). The presence of albumin in urine is a well-established biomarker for the early detection of diabetic nephropathy. We have developed a technique for electrochemical detection of microalbuminuria for point of care applications by exploring the binding property of human albumin with electrochemically active molecules like copper and hemin. Methylene blue mediated sensing technique has also been proposed. Urine Albumin-to creatinine ratio (ACR) is another variant of the microalbumuria test that can be done any time and does not suffer from the dilution factor of urine. Iron binding property of creatinine is exploited to develop creatinine biosensor, thus enabling POC ACR tests.

L’anémie est fréquente chez les patients pédiatriques en postopératoire de chirurgie cardiaque. Malgré le nombre important de patients transfusés, le taux d’hémoglobine (Hb) pour lequel les bénéfices surpassent les risques est inconnu chez ces patients. Récemment, Lacroix et al. ont démontré qu’une stratégie transfusionnelle restrictive n’était pas inférieure à une stratégie libérale en ce qui concerne le développement ou la progression du syndrome de défaillance multiviscérale (SDMV) et la mortalité chez les patients de soins intensifs pédiatriques (SIP).Devant le manque d’évidence, une analyse de sous-groupes des patients en postopératoire de chirurgie cardiaque de l’étude Transfusion Requirements in Pediatric Intensive Care (TRIPICU) a été réalisée. L’objectif de cette étude était de déterminer l’impact d’une stratégie transfusionnelle restrictive comparée à une stratégie libérale sur l’acquisition ou l’aggravation du syndrome de défaillance multiviscérale (SDMV) chez les enfants en postopératoire de chirurgie cardiaque. Cette étude n’a pas démontré de différences statistiquement, ni cliniquement significatives du nombre de patients ayant acquis ou aggravés un SDMV, ni des issues secondaires entre les stratégies transfusionnelles restrictive et libérale. L’analyse de sous-groupes permet de générer une hypothèse de recherche et les résultats devraient être confirmés par un essai randomisé contrôlé. / Anemia is frequent in pediatric patients following cardiac surgery. Despite frequent transfusions, the optimal hemoglobin threshold where benefits surpass risks is still unknown for these patients. Recently, Lacroix et al. showed that a restrictive transfusion strategy was not inferior to a liberal strategy concerning the development or progression of multiple organ dysfunction syndrome (MODS) and mortality in pediatric intensive care patients. In the absence of evidence, the aim of this study was to determine the impact of a restrictive versus a liberal transfusion strategy on new or progressive multiple organ dysfunction syndrome (MODS) in children following cardiac surgery. We conducted a subgroup analysis of the postoperative cardiac surgery patients of the Transfusion Requirements in Pediatric Intensive Care Unit (TRIPICU) study. Our study showed no statistically and clinically significant differences in the number of patients who acquired or worsened MODS, nor secondary outcomes between a restrictive and a liberal transfusion strategy. This subgroup analysis generates a research hypothesis that should be confirmed by a randomized controlled trial.