Exam 2 Guoqing Wang 1. Metabolomic analysis of changes to plasma metabolome following dietary depletion and supplementation of thiamine pyrophosphate (TPP) in mice Experimental design Male C57BL/6J mice (No. 11401300006402), weighing 20 ± 2 g, will be used for this experiment. 7-month old mice will be maintained in a room at 23 ± 2°C and a relative humidity of 50 ± 10%, with a natural light-dark cycle. Food (6 mg thiamine-HCl as the form of TPP) and water will be provided ad libitum. After acclimatization for 1 week, the mice will be divided into two groups (n = 10 in each group) based on dietary difference: the dietary TPP depletion group (no TPP added) and the dietary TPP supplementation group (6 mg thiamine-HCl/kg diet as the form of TPP). The supplementation level will be based on Nutrient Requirements of Laboratory Animals: Fourth Revised Edition, 1995 [1]. Plasma samples will be collected from mice in each group before feeding the experimental diets and after 10-day feeding period, the length of feeding experimental diets will be based on a previous publication [2]. Approach Plasma samples will be analyzed by H-NMR with or without filtration and by targeted quantitative by mass spectrometry (MS) according to Gregory III et al., 2013 [3]. NMR spectral will feature selected metabolites including acetyl-CoA, succinyl-CoA, acetoacetate and ribose. Expected results Compared to dietary thiamine supplementation at the recommended level, thiamine depletion will decrease
In the scientific method lab, I explored the different food options for mice in order to find the best one that will help them gain weight. To succeed in this lab, I needed to examine several types of foods including fruits, raw meat, cooked and uncooked foods. In this experiment I used cooked macaroni as a positive control and for the negative control I didn’t feed the mice any food, only water. The main objective of this study was to determine the best solution that will help me understand the metabolism of mice. All mice used in this study were given the same resources, but the only difference was the different types of foods fed to the mice to understand the changes of weight gain or loss between the mice.
Male AP+ Tg Sprague-Dawley (SD) rats weighing 350-450 g will be used in this study. There will be a total of 40 rats which will be divided into four groups with ten in each group. Adult DRG from C1 to L1 will be dissected from rats ≥ 8 weeks of age using standard techniques.
A total of 840 chick (Ross) one day old will be separated into 7 treatments. The first group is control (basal diet), the second is phosphorus 5g (basal diet + 5g / kg diet), the third is phosphorous 1g (basal diet + 1g / kg diet), the fourth is phytase 500 FTU (basal diet + 500 FTU / kg diet), the fifth is phytase1500 FTU (basal diet + 1500 FTU / kg diet), the sixth is probiotic 0.5 g (basal diet + 0.5 g / kg diet),
Male Wistar rats between 120 and 140 kg will be fed an ammonium acetate rich diet to induce them with hyperammonemia for a 5-week period. Thirty Wistar rats will undergo surgery to implant portacaval shunts. This model was created to develop a system that has a pure hyperammonemic state that is commonly found in individuals with chronic liver disease. Four groups of 10 Wistar rats, split into 2 control groups and 2 experimental groups. The four designated groups will be: the normal fed control group, the control + PMO group, the portacaval shunted group (ammonia group), and the portacaval shunted Wistar rats + PMO groups. Portacaval shunts will be performed, in order to induce alterations in nitrogen metabolism caused by changing the function of the liver. The control + PMO group and portacaval shunted Wistar rats + PMO group will be given 60 mg/L of PMO in their drinking water everyday at the same time until their deaths. In order to ensure all water will be consumed, all Wistar rats will be provided with only 10 ml of water. The Wistar rats in the control groups will be given tap water on the same drinking schedule as the two experimental groups. After 5 days of treating each experimental group, the Wistar rats willingness to explore brain function was assessed using a Y-Maze learning test everyday for 10 trials. Additionally, ammonium
The results for the baseline metabolic rate were as expected. The metabolic rates of the Tx and the Hypox rat were lower than the metabolic rate of the normal rat. The Tx rat could not produce thyroxine because it had no thyroid gland. The Hypox rat could not produce TSH which stimulates the thyroid to produce thyroxin.
THs exert a wide series of effects acting upon virtually all tissues in the organism( Venditti ,et al.,2003). The actions derived from the THs are not well known and seem to differ significantly. For instance, diiodothyronin (T2) produces metabolic effects similar to those of T3 (Lanni,et al.,1996) whereas thyronamines oppose its actions (Venditti, et al.,2011), at least at the mitochondrial level .The known actions of the THs can be grossly classified in two general processes: regulation of growth and development, and metabolism regulation. The metabolic effects of THs are
Figure 3 shows the percentage of gain in body mass compared to the initial mass of the body at the 6, 12, and 18-week time points. The vitamin D deficient groups showed the lowest percentage of body mass gain compared to the vitamin fed groups. Even though smoking was independent, and played no affect in body mass, the smoking vitamin D deficient mouse showed the lowest body mass gain at every single point.
The different groups consisted of MK-801 injections, Saline injections and no injections. The injections were given twice daily for a week long period. The animals were then tested twice daily for a three week long period using 2 methods. The methods consisted of an experimental paradigm and a control paradigm. The experimental paradigm released a pellet that consisted of 45mg dustless precision food pellet every minute for the two hours of testing (120 minutes/2hr = 120 pellets). The control paradigm provided the 120 pellets freely in a dish. Both groups had free and equal access to water throughout the experiment. They had measured which group had consumed more water by weighing the water bottles before and after the
ATP is an essential molecule for the body to function and is found in all cells. It is used for energy production and transportation, and as a result, without ATP, the body would cease to function sufficiently. The amount of ATP produced and used by the body can be determined by the oxygen consumption rate, so a higher oxygen consumption rate will indicate a higher metabolic rate (Storey and Storey, 2003; Ross, 2006). Animals that have their diet restricted or are in a state of starvation have shown a decline in metabolic rates as well as a difference in behaviour. Previous studies have shown that when an animal is starved it is more likely to undertake behaviour of extreme risk to acquire food, leading to a lower survival rate in animals in this category (Hazlett 2003; McCue 2010; Bohrer & Lampert 1988). Young animals that are not on an adequate nutritional diet will not maintain a standard growth rate. Females may not be able to undertake reproduction or reach a reproductive stage due to the increased need of energy and nutrition during this process, and if reproduction is undertaken, there will be a high chance of detrimental effects to both the mature animal and offspring. (Marsden, et al.
The animal experiment was approved by the University of Calgary Animal Care Committee (#AC12–0033). 32 male Sprague Dawley rats were ordered when they were 10 days old (Charles River, Montreal, QC, Canada) and randomized into treatment groups after a two-week adaptation interval. The four treatment groups were as follows (n=8): 1. Control (CON, animals fed ad libitum diet) 2. 75% restriction group (75%, animals fed 75% of the diet consumed by the control animals) 3. 50% restriction group (50%, animals fed 50% of the diet consumed by the control animals) 4. 50% restriction group and then switched to ad libitum diet after two weeks (50% +CON). The rats weighted ~80g when housed individually in metabolic cages of the Comprehensive Lab Animal Monitoring
The rats in the HFD groups fed a high fat diet [29% fat plus 1% cholesterol (Merck Germany), 0.5%cholic acid (sigma USA), 23% protein, 38% carbohydrate, 5/5% ash, 3% fibers] with 4700 Kcal ME/kg. ME content of diets was calculated based on pet food manufacture’s associate propose.
Untargeted metabolomics profiling of liver and brain samples from SD mice (n=3) and normal mice (n=3) were performed through RPLC. A total of 177 metabolites were found to be significantly dysregulated in mouse liver samples (p (-- removed HTML --) 0.5). Out of these 177 metabolites, 96 (54.2%) out were significantly upregulated, while 81 (45.8%) were downregulated in SD mouse liver. Similarly, a total of 112 metabolites were found to be dysregulated in mouse brain samples (p (-- removed HTML --) 0.5). Out of these 112 metabolites, 53 (47.3%) were significantly upregulated, while 59 (52.7%) were downregulated in SD mouse brain. Further, the same technology was
Control (n=3 rats): Rats received rat chow and tetracycline (500mg/liter) in their water for 8 days, starting from 21 days of age to 29 days of age.
The subjects were Sprague-Dawley male rats. Their age is 150 days. The supplier is Harlan Sprague-Dawley. They are maintained on a 12:12 h light/dark cycle and are provided with ad libitum access to food.
Male Sprague-Dawley (SD) rats (weighing 280–300g) (n=90) ) from the animal laboratory of Shiraz University of Medical Sciences, Shiraz, were used for the experiments. Animals were provided standard laboratory rodent food and water ad libitum and were housed in a temperature-controlled environment of 23±2oC and a relative humidity of 45±10% on a 12-h light/12-h dark cycle. All animals did not get additional exercise and were allowed for normal activities in a cage with dimensions of 65×45×30 cm. There is no exercise protocol during the study period. Animal experiments were performed in accordance with the internationally accredited guidelines, and have been approved by the author’s institutional Animal Care and Use