In Table 1 I have my substance of FAS,SREBP-1 and Actin and those are what I places in the wells of the gel to receive my results that I received in figure in Figure 5 picture F. If you compare me and my classmates primer optimization results, you can see that we all have a spear of the ladder on our gel. Another similarity is that we all did receive bands for FAS, SREBP and Actin and our cDNA came out very good with a dark band. Figure 6 is a repeat of Figure 5 picture F and the reason for repeating this experiment was because we were trying to see what other temperatures would work for Actin. Figure 7 is a western blot of SREBP-1c and it came out pretty nice. My cDNA was visible and the licor and Kal showed up very good and dark. My classmate actually performed the CPT-1 (Figure 8) western blot and you can barely see any bands for the cDNA but the actin and CPT-1 is very visible and it came out very dark. For the FAS (Figure 9) the licor and kal showed up very nice and bight red and there are bands for the cDNA visible and the Actin is visible as well. Fort he PPARα western blot, we have beautiful bands for the Cdna, Actin and the kal. If you compare Figures 7-10, you can see a resemblance in the kal, licor and Actin showing up better than the cDNA. If the cDNA is not showing up that well, maybe we need to increase the amounts of cDNA that we place in the wells. Figue 11 shows the PPARα western blot and Qpcr. For the western blot, there is no significant difference of protein PPARα but for the qPCR, the analysis shows that there is a significant increase in the IUGR compared to the control at approximately at 3 folds. For figure 12 western blot of PEPCK, there is a significant increase in the IUGR versus the control. For the PEP...
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4. White, Brett, MD. Dietary Fatty Acids. 8/2013. http://www.aafp.org/afp/2009/0815/p345.html. Los Angelas, CA. 80(4):345-350.
5. Brandt, Mark, Ph.D. Introduction to Lipid Metabolism. 2000-2003. http://www.rose-hulman.edu/~brandt/Chem330/Lipid_metab.pdf
6. Claudio Souza, Marisa Frederico, Gabriella Luz, Dennys Cintra, Eduardo Ropelle, Jose Pauli and Licio Vellosos. Acute exercise reduces hepatic glucose production through inhibition of the Foxo1/HNF-4α pathway in insulin resistant mice. 5/2010. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2911223/. 588(Pt12)
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7. No Author. Liver Function, Facts, Anatomy, and Disease. Web.2/2014. .
8. No Author. Metabolic Functions of the Liver. Web. 4/2014. http://www.vivo.colostate.edu/hbooks/pathphys/digestion/liver/metabolic.html.
9. Excel Graphing/Charting
Digestion of the haemolytic and non-haemolytic cells allowed for easier identification of fragments during electrophoresis analysis. Lane 12 in figure 3 show the size markers of SPP1 digested with EcoR1 while lanes 6 and 7 show samples of pK184hlyA and pBluescript digested with EcoR1 and Pst1. Lane 4 was loaded with plasmid DNA from haemolytic cells digested with EcoR1 and Pst1 while lane 5 was loaded with EcoR1 and Pst1 digested DNA from non-haemolytic cells. There was a lack of technical success in both lanes due to no bands appearing in lane 4 and only a single band appearing in lane 5. Theoretically, two bands should appear in both lanes after successful to allow for fragment identification. A possible explanation for the single, large fragment in lane 5 is that successful digestion did not take place and the plasmid was only cut at one restriction site leaving a large linear fragment of plasmid DNA. The absence of bands in lane 4 could be because there was not enough plasmid loaded into the lane. Another possibility could be that low plasmid yield as obtained when eluting the experimental samples in order to purify it. Lanes 8 and 9 belonged to another group and show technical success as two bands were present in both the haemolytic (lane 8) and non-haemolytic (lane 9) lanes. If the
Thyroid and metabolism hormones play a large role in the daily lives of all living species. Thyroid hormones regulate the metabolism and the metabolism is responsible for maintaining a specific range for the biochemical reactions that occur in the body (Martini 2014). The most important hormone for metabolic maintenance is thyroxine (T4). This hormone also plays a large role in body heat regulation. It is produced by the pituitary gland and secreted by the thyroid gland. The thyroid releasing hormone (TRH) must trigger the thyroid stimulating hormones (TSH) to release thyroid hormones to the thyroid gland. These hormones are under control of the hypothalamus, or main neural control center. Propylthiouracil (PTU) is a medication used to treat
time to mix the reaction solution after adding the Taq polymerase. When the PCR technique is
EDTA, the chelating agent that binds with magnesium, had a high absorbency and strong color change to red. The correct cofactor was copper which with the chelating agent of PTU and citric acid which both bind strongly to copper which keeps it from binding with the enzyme. This was determined because in the trails, both PTU and citric acid had low absorbency and were clear or roughly clear in color. The catechol in each tube, which was the control for this experiment, allowed the cofactor that would be used in this reaction to be singled out. The way each chelating agent would affect the different cofactors displayed which was not needed for the reaction and which cofactors were needed for the reaction. An inconsistency that may have affected the data would be if the calibration tube malfunctioned in balancing the spectrophotometer to zero. There also could be errors if the calibration tube wasn’t used before each tube was tested in the spectrophotometer. The relationship of the cofactor and amount of enzyme activity would be that if the cofactor is inhibited or not, the enzyme activity would be higher if the cofactor is not inhibited but lower if it was inhibited by the chelating
Insulin is a key player in the control of intermediary metabolism. It has profound effects
The pathophysiology of type 2 diabetes is characterized by both insulin resistance and insulin secretion. Peripheral insulin resistance and inadequate insulin secretion by the pancreas due is due to beta cell dysfunction. The resistance results in elevated free fatty acids and inflammation which leads to an overall decreased amount of glucose taken up into the muscle, increased glucose production, and incr...
9. Zaharieva, D. P., & Riddell, M. C. (2013). Caffeine and glucose homeostasis during rest and exercise in diabetes mellitus. Applied Physiology, Nutrition & Metabolism, 38(8), 813-822.
Glycemic index is the measure of how quickly blood glucose rise after eating a particular kind of food. This is used by estimating the how much each gram of carbohydrate consumed raises a person’s glucose level. During intense exercise, the body uses glucose as energy source before it starts relying of fats. “Prolonged exercise can only be continued when there is an adequate amount of carbohydrate available to fuel muscle and the brain”(William,2004). This shows that the amount of carbohydrate/glucose that is found within the body’s tissue has a significant amount of influence to play on how effective the early hours of an exercise would be. Glycemic index helps one to know how effective the carbohydrate/glucose within the body is functioning during exercise.
...ll cause your blood sugars to lower (Mayo Clinic Staff). While working out you will notice that your blood sugar levels will be all over the place so its important to have things such as glucose pills just in case your levels drop too low. Exercise may not be the cure for diabetes but it could be the thing that prevents you from getting it.
Heilbronn, Leonie K., and Campbell, Lesley V.. “Adipose tissue macrophages, low grade inflammation and insulin resistance in human obesity.” Curr Pharm Des 2008, 14:1225-1230.
We used the Sniffy the virtual rat Pro Version 3.0 program by Wadsworth Cengage Learning. This program attempts to accurately simulate some of the psychological processes and behavioral phenomena found in rats. In this program, all of the subjects are always starved for food, regardless of how much it has eaten. This simplifies the process of operant conditioning by reducing the amount of time required to simply wait for the virtual rat to be hungry again.
Setting up the apparatus was quick and easy, but measuring the DCPIP solution took a bit of time because they had to be the same volume so that they don’t affect the results. Although there were variables that were controlled, there was another variable which I did not identify until after our investigations were done.
Gel electrophoresis is used in a variety of settings, particularly in molecular biology. Besides being used to separate nucleic acids, such as DNA and RNA, gel electrophoresis is also employed to divide proteins (Gel Electrophoresis). According to research, electrophoresis is applied for the following reasons, "To get a DNA fingerprint for forensic pur...
The first step was to obtain the White Rat and to tie it in the supine position, anterior surface facing up in side the dissection pan. To tie the animal, we used butcher’s twine and secured the front and hinds legs using a “lasso” technique, careful not touch the sharp claws. To make the first insicion I had to locate the Xifoid Process of the rat (distal aspect of the sternum). Once I had located the Xifoid Process, I had to use forceps to pull the skin of the animal’s abdomen up and use the scissors to cut. The first incision is made from stem to sternum, cutting through the errectos abdomen muscle down to the groin. The second incision ion is perpendicular to the first below the diaphragm. Because of this technique we were able to open the abdominal cavity first. The third and forth incisions were made bilaterally above the legs. The last two incisions were made in upside down “V” shape on the collarbone, to expose the thoracic cavity. This dissection was both sharp, because of the use of the scissors and scapel and blunt because of the use of the probe and forceps to move organs and skin to expose other organs not yet identified.
...tochondrial lipid uptake (6). Therefore, evidence that lipid oxidation decreases throughout the body in people with obesity and insulin resistance as a result of impaired mitochondrial plasticity (7). It still remains unknown if insulin has direct and rapid effects on mitochondrial function. If insulin is responsible, the defects caused by IR might reflect deregulation of the lipid‑induced PPAR–PGC1 interaction after prolonged hyperlipidemia (8). This in effect leads to reduced lipid uptake into mitochondria in order to balance for lower mitochondrial content and increased lipids concentrations. As a result of this reduced lipid uptake, lipid induced uncoupling of the respiratory chain takes place, there is reduced oxidation of glycolytic substrates, which will detach fatty acid oxidation rates from citric acid cycle rates, resulting in metabolic distortion (7).