BoD Lipid Peroxidation Report

BoD Lipid Peroxidation Report A Study of lipid peroxidation The degradative process of lipid peroxidation in the liver and the potential of antioxidants to prevent cell damage Lipid peroxidation of rat homogenate using the Fenton reaction to generate free radicals (-OH and -O2) to initiate the self-propagating peroxidation of cell membrane fatty acids. Two separate antioxidants were used (aTocopherol and Quercetin) to study the potential of antioxidants in the prevention of cell damaged. Data of two separate groups (A+B) was provided along with data enabling the construction of a calibration curve to measure local MDA concentrations as an indication of peroxidation damage. The Fenton reaction produced the highest concentration of MDA in both data sets which is expected, allowing for a comparison of free radical damage in the presence of antioxidants. In the presence of aTocopherol, there was an MDA (nM/ml) concentration reduction from 45nM/ml to 24nM/ml evidencing a peroxidation inhibition via the binding of free radicals to the antioxidant though some damage was still caused as MDA concentration was higher than the control (7nM/ml). Quercetin showed a com plete reduction in local MDA concentration from 68nM/ml to 7nM/ml, which is equal to that of the control; evidencing a complete lipid peroxidation via the binding of all free radicals produced and thus prevents cell damage. Lipid peroxidation is the multistep process of oxidative degeneration of lipids. The process involved polyunsaturated fatty acids and the free radicals -OH (hydroxide) and -O2 (superoxide), which are unstable forms of oxygen to the incomplete valence ring on their outer shell resulting in an unpaired electron (free electrons). Due to the naturally unstable state of a single unpaired electron, free electrons are highly reactive (free radicals) requiring an electron to become stable; making the unpaired hydrogen atoms on the fatty acid tails suitable for binding (Mylonas C, 1999). The three step process (initiation, propagation and termination) of lipid degenerative produces highly reactive electrophilic aldehydes, which react with CH2 group forming CH (carbon centred) radicals. CH radical then reacts with O2 radicals producing peroxyl radicals (Yngo J. Garciaa, 2005). This propagation reaction then reacts with adjacent CH2 groups resulting in the formation of lipid hydroperoxide. Lipids are essential components of cell membranes (i.e. phospholipids and glycolipids) and can be used in the identification of damage as a result of the pathogenesis of disease via reactive oxygen species (ROS) concentration. ROS-dependent tissue damage can be identified by increased local MDA (malonedialdehyde) and 4-HNE (4-hydroxynonenal) (Kwiecien S, 2014). MDA is the product of lipid peroxides metabolisation, and can be indicative of oxidative stress related disease i.e. atherosclerosis, and induced gastric injury (due to gastric mucosa damage). Due to free radicals are reactive its uncommon that they a found in that state as they tend to bond and react very quickly in order to fill their valence shell and become stable. The Fenton Reaction (Fe2+ and H2O2) issued to generate free radicals (particularly -OH) and initiates lipid peroxidation within the liver. During the breakdown of lipids, malonedialdehyde (the final product of lipid breakdown) reacts with thiobarbituric acid resulting in a testable pink adduct. The Fenton reaction is as follows: Fe+2 + H2O2 > OH (hydroxyl ion) (Fenton Reaction) OH + lipid > malonedialdehyde Malonedialdehyde + thiobarbituric acid > thiobarbituric acid reactive substance (pink) Set up a series of test tubes a labelled and the volumes laid out in Table 1 were pipetted into the corresponding tubes. Remember to add the rat homogenate last due to this starting the reaction. The tubes were then incubated for 30 minutes at 37 degrees Celsius. At this point, the standard curve of MDA was set up as seen in Table 2 and tested at a wavelength of 532nm. After which thiobarbituric acid was added to the original test tube set and incubated for a further 15 minutes in after the adduct fluid was removed and tested at 532nm. Test Tube Test Buffer Tris HCL (00.2M) pH 7.2 FeCl2 H2O2 Catalase Quercetin OR aTocopherol Homogenate Total/ml 1 Control 1.6ml 0.9ml 2.5 2 Fe2+ 1.1ml 0.5ml 0.9ml 2.5 3 Fe2+/H2o2 0.6ml 0.5ml 0.5ml 0.9ml 2.5 4 Catalase/Fe2+/H2o2 0.5ml 0.5ml 0.5ml 0.1ml 0.9ml 2.5 5 aTocopherol or Quercetin /Fe2+/H2o2 0.5ml 0.5ml 0.5ml 0.1 0.9ml 2.5 Table 1: test tube volumes for each of the five test tubes in the lipid peroxidation assay, empty spaces indicated that the solution isnt added to that tube. Each was incubated for 30 minutes together under the same conditions. Test Tube Final MDA concentration (mM) Dilutions Volume of MDA stock (ml) Buffer (ml) Total Volume (ml) 1 0.1 Dilute 1mM MDA 1:10 0.3 2.7 3 2 0.05 Dilute 0.1mM MDA 1:2 (tube 1 extract) 1.0 1.0 3 3 0.01 Dilute 0.05 mM MDA 1:5 (tube 2 extract) 0.4 1.6 3 Table 2: The dilutions volumes of MDA and the final concentration required, these volumes were used to construct a calibration curve for comparison of the test samples in table 1. NOTE: all data using in the results was provided, this was due to an issue in the lab were where independent data was unintentionally taken by another individual and thus leaving no results for comparison against overall class data. MDA Concentration (nMoles/ml) Optical Density (OD) at 532nM 0 0 12.5 0.07 25 0.145 50 0.26 100 0.55 Table 3: MDA concentration (nMoles/ml), these values were used to construct the calibration curve Figure 1. MDA concentrations were provided due to an issue with both groups overall dilution series. The data from figure 1 was plotted using table 3. The R2 value (0.9986) indicates a strong linear value between the MDA concentrations (nM/ml) and the optical density. Figure 1: A calibration curve using the data from Table 3. The data set shows a strong linear relationship between optical density and known MDA concentration indicating good lab practice. Tube Mean -/+ Stdev SEM 1 Control 0.068 0.077 0.063 0.006 0.073 0.045 0.074 0.058 -/+ 0.025 0.010 2 Fe2+ 0.082 0.081 0.057 0.03 0.003 0.050 0.075 0.054 -/+ 0.029 0.011 3 Fe2+/H2o2 0.174 0.247 0.093 0.577 0.058 0.319 0.251 0.246 -/+ 0.173 0.065 4 Catalase/Fe2+/H2o2 0.355 0.169 0.246 0.063 0.056 0.143 0.134 0.167 -/+ 0.105 0.040 5 aTocopherol/Fe2+/H2o2 0.074 0.173 0.074 0.127 0.259 0.092 0.110 0.130 -/+ 0.666 0.025 Table 4: class data group A using aTocopherol, the values were done in repeat to gain a mean value and allows for Stdev calculation and thus SEM calculation, allowing for later comparison. The data set in Table 4 was provided and used the antioxidant aTocopherol. Seven repeats of each test were conducted to allow for a mean to be gained and thus a Stdev and then a standard error mean. The error mean allows for comparisons between different data sets as it indicates how accurate the experiment was rather than how varied (Stdev). The data was plotted in figure 2 and 3 with the variation of containing either the Stdev (figure 2) or the SEM (figure 3). Figure 2 allows for variation comparison while figure 3 allows for accuracy comparison between the two data sets (group A and Group B). Figure 2: the mean OD values of aTocopherol, the error bars show the variation within the data set. Test tube 2 was the most optically dense of the data set while test tube 2 was the least, though the error bar would suggest some variation in this value considering test tube 1 (control) was more optically dense. Figure 2 shows the optical density of aTocopherol. Test tube 1 contained only buffer and showed little variation between repeats resulting in a small Stdev, while test tube 4 has a large Stdev value and thus would need repeating in order to gain an accurate representation of the data. Test tube 3 was the most optically dense with a value 0.246 (at 532nm), while the OD went down between test tubes 4-5 (0.167 and 0.130). This is visually shown in in figure 3, where the data was plotted in a bar graph and SEM was used to show the accuracy of the experiment. The deviation of the error bars shows high accuracy in some results i.e. test tube 1-2-3. However, the deviation in test tubes 4-5 was high compared to other samples. Figure 3: the graph shows the class data of group A. The mean OD values of aTocopherol were plotted including the SEM to show how accurate the experiment was between data sets. Test tube 3 showed to be the most optically dense of the set while test tube 2 showed to be the least.   Ã‚   Tube Mean -/+ Stdev MDA concentration (nM/ml) 1 Control 0.058 -/+ 0.025 7 2 Fe2+ 0.054 -/+ 0.029 7 3 Fe2+/H2o2 0.246 -/+ 0.173 45 4 Catalase/Fe2+/H2o2 0.167 -/+ 0.105 28 5 aTocopherol/Fe2+/H2o2 0.130 -/+ 0.666 24 Table 5: a table showing the MDA concentrations of Group A class data set of each test tube using the calibration curve in Figure 1. Table 5 shows the MDA concentration of group A using aTocopherol, the control had the sample concentration of MDA as the Fenton reagent (7nm/ml); while test tube three which contained the Fenton reagent and H2O2 resulted in the highest MDA concentration of (45nM/ml). Adding the antioxidant resulted in a reduced MDA concentration of 24nM/ml. The visualisation of Table 5 data is seen in Figure 4 where MDA concentration is plotted against each test tube value (gained from the calibration curve) Figure 4: The graph shows the MDA concentration (nM/ml) of the groups A class data set, as only one set of samples was done no comparison can be made between the same antioxidant via Stdev. Test tube 3 showed to contain the highest concentration of MDA (45nM) while test tube 2 also showed to contain the lowest concentration of MDA (7nM). Tube Mean -/+ Stdev SEM 1 Control 0.041 0.06 0.08 0.057 0.057 0.02 0.297 0.087 -/+ 0.094 0.036 2 Fe2+ 0.037 0.039 0.06 0.06 0.053 0.074 0.047 0.053 -/+ 0.013 0.005 3 Fe2+/H2o2 0.28 0.704 0.242 0.365 0.247 0.385 0.528 0.393 -/+ 0.170 0.064 4 Catalase/Fe2+/H2o2 0.14 0.497 0.087 0.305 0.351 0.099 0.357 0.263 -/+ 0.156 0.059 5 Quercetin/Fe2+/H2o2 0.046 0.035 0.035 0.073 0.073 0.031 0.102 0.056 -/+ 0.027 0.010 Table 6: The table shows the class data set of group B using Quercetin as an antioxidant, multiple repeats were undertaken to allow for an average to be gained and Stdev and SEM to be calculated. The control only contained buffer solution. Figure 5: The graph shows the mean OD of the group B class data set, using quercetin as an antioxidant. Stdev values were used as error bars to visualise the variation between the dataset. Test tube 3 showed to be the most optically dense while test tube 2 showed to be the least though showed high Stdev and thus a lot of variation between the individual repeats. Figure 6: The graph shows the mean OD of the group B class data set, using quercetin as an antioxidant. SEM values were used as error bars to visualise the variation between the dataset. Test tube 3 showed to be the most optically dense while test tube 2 showed to be the least though showed high SEM and thus low accuracy between the individual repeats. Tube Mean -/+ Stdev MDA concentration (nM/ml) 1 Control 0.087 -/+ 0.094 15 2 Fe2+ 0.053 -/+ 0.013 7 3 Fe2+/H2o2 0.393 -/+ 0.170 68 4 Catalase/Fe2+/H2o2 0.263 -/+ 0.156 46 5 Quercetin/Fe2+/H2o2 0.056 -/+ 0.027 7 Table 7: a table showing the MDA concentrations (nM/ml) of Group b class data set of each test tube using the calibration curve in Figure 1. Table 7 shows the MDA concentration of group B using quercetin, the control had the sample concentration of MDA as the Fenton reagent (15nm/ml); while test tube three which contained the Fenton reagent and H2O2 resulted in the highest MDA concentration of (68nM/ml). Adding the antioxidant resulted in a reduced MDA concentration of 7nM/ml. The visualisation of Table 7 data is seen in Figure 7 where MDA concentration is plotted against each test tube value (gained from the calibration curve) Figure 7: The graph shows the MDA concentration (nM/ml) of the groups B class data set, as only one set of samples was done no comparison can be made between the same antioxidant via Stdev. Test tube 3 showed to contain the highest concentration of MDA (68nM) while test tube 2+5 also showed to contain the lowest concentration of MDA (7nM). NOTE: Due to individual data being lost only a comparison between the two data class data set can be made The enzymatic destruction (via catalase, superoxide dismutase) of membrane lipids is a crucial step in the pathogenesis of multiple disease states within adult (Mylonas C, 1999), the reactive oxygen species (hydrogen peroxide) produced during lipid peroxidation readily attacks the polyunsaturated fatty acids within the phospholipid bilayer causing the commencement of a self-propagating chain reaction within the membrane due to CH radicals reacting with O2 radicals producing peroxyl radicals (AW, 1998). Due to the self-propagating nature of the reaction series small lipid peroxidation can cause serious tissue damage resulting in atherosclerosis, asthma or kidney disease. Antioxidant activity quenches molecular oxygen (Yamauchi, 2010), and helps in the stabilisation of lipid-peroxyl free radicals via inhibition. Quercetin, a plant-derived aglycone flavonoid (Zhang M, 2011) was compared to aTocopherol (vitamin E) in the lipid peroxidation of rat liver homogenate. The liver metabolises materials and thus results in the production of free radicals when the oxidative balance is lost it leads to oxidative stress and thus having antioxidants to restore homoeostasis is required. Antioxidants have a high affinity for free radicals (Muriel, 2015) due to their ability to donate electrons. The antioxidant a-Tocopherol reduces oxidation under strong oxidative conditions, reducing the number of free radicals to be free at the end of lipid peroxidation. The data in figure 2 shows the average OD including Stdev bars, the variation in tubes 4-5 indicates poor experimental practice resulting in poor repeats within the data set and thus increasing variation within the data set. It suggests high oxidative conditions in tube 3 producing high concentrations of MDA (nM/ml) as seen in figure 4. Figure 4 also evidences that in the presence of a-Tocopherol lipid peroxidation is reduced as a reduction of MDA (the final product of lipid peroxidation and would result in pink adduct) is being produced suggest an interruption in the self-probating cycle of the fatty acids within the liver homogenate. This reduction is evidence as MDA concentration goes from a peak of 45nM/ml to an MDA reduction 24nM/ml in the presence of a-Tocopherol. When comparing the two sets of Data SEM and SD is used in order to give a relative comparison between the two different groups due to them being undertaken under different conditions. Comparing figure 2 and figure 5 (which used SD) the variation in data set A was much more significant as the higher SD values indicating a large variation within the repeats evidencing low reliability. Figure 5s SD bars a smaller then figure 2 indicating less variation and an increased reliability of the obtain results. Though both sets of data (A-B) show that the highest OD was found to be within tube 3 indicating that Fe2+ and H2O2 produce the highest concentration of MDA (nM/ml). SEM of the two data sets show that the accuracy of the two groups are similar and both show a decline in MDA concentration in the presence of the antioxidant, evidencing a reduction in lipid peroxidation (MDA is the product of lipid peroxides metabolisation which results in the pink adduct) and free radical production in the presence of the chosen antioxidants. Using the calibration curve to gain the MDA concentration of each antioxidant shows that quercetin resulted in a total reduction of free radicals as the MDA concentration was reduced to that of the control (buffer solution). Comparing this to a-Tocopherol there was a reduction of nearly half free radical concentration. These results indicate that the levels of oxidative stress are reduced in the presence of antioxidants. Improvements that can be made include, not losing the individual samples which would have been used for comparison, increasing the amount of antioxidants used to show and overall reduction in free radicals in different antioxidants. Also individual human error resulted in data sets begin provided requiring more lab expertise would reduce this and thus reduce was and cost of the experiment. Antioxidants reduce the concentration of MDA (nM/ml) present in the test tube via the inhibition of oxidative stress and lipid peroxidation of the cell membrane lipids. Quercetin completely reduced local MDA concentration of the rat homogenate indication no lipid peroxidation was occurring due to the binding of antioxidant to the local free radicals (produced via the Fenton reaction) due to their naturally high affinity. There was also a noticeable reduction of MDA concentration in the presence of aTocopherol though this was only an estimated 50% reduction. It can be seen that antioxidants offer a level of cell lipid protection against free radicals and a reduction in oxidative stress, resulting in less overall tissue damage. References Antonio Ayala, M. F. (2014). Lipid Peroxidation: Production, Metabolism, and Signaling Mechanisms of Malondialdehyde and 4-Hydroxy-2-Nonenal. Oxidative Medicine and Cellular Longevity, 2014(2014), 31. doi:http://dx.doi.org/10.1155/2014/360438 AW, G. (1998). Lipid hydroperoxide generation, turnover, and effector action in biological systems. The Journal of Lipid Research, 39(8), 1529-1542. Esterbauer H, G. J. (1992). The role of lipid peroxidation and antioxidants in oxidative modification of LDL. Free Radical Biology and Medicine, 13(4), 341-390. Justino GC, S. M. (2004). Plasma quercetin metabolites: structure-antioxidant activity relationships. Archives of Biochemistry and BIophysics `, 432(1), 109-121. doi:10.1016/j.abb.2004.09.007 Kwiecien S, J. K. (2014). Lipid peroxidation, reactive oxygen species and antioxidative factors in the pathogenesis of gastric mucosal lesions and mechanism of protection against oxidative stress induced gastric injury. Journal of Physiology and Pharmacology, 65(5), 613-622. Muriel, S. C.-G. (2015). Antioxidants in liver health. The World Journal of Gastrointestinal Pharmacology and Therapeutics, 6(3), 59-72. doi:10.4292/wjgpt.v6.i3.59 Mylonas C, K. D. (1999). Lipid peroxidation and tissue damage. In Vivo, 13(3), 295-309. Yamauchi, R. (2010). Functions of Antioxidant Vitamins against Lipid Peroxidation. (F. o. Science, Ed.) Foods Food Ingredients Japan, 215(1), 501-1193. Yngo J. Garciaa, A. J.-M. (2005). Lipid peroxidation measurement by thiobarbituric acid assay in rat cerebellar slices. Journal of Neuroscience Methods, 144(1), 127-135. Zhang M, S. S. (2011). Antioxidant properties of quercetin. Advances in Experimental Medicine and Biology, 701, 283-289. doi:doi: 10.1007/978-1-4419-7756-4_38.

Ensuring Freedom by Preserving the Values of Trade Unions Essay

Ensuring Freedom by Preserving the Values of Trade Unions Thoughtful committed citizens are the only thing that have ever changed the world. —Margaret Mead Anti-union sentiment is increasingly pervading American culture. In fact, one critic says, â€Å"The United States in now on the verge of a risky experiment: to become the first parliamentary democracy in modern world history without a substantial trade union movement† (Lichtenstein 66). In addition to weakening bargaining power, the judicial system allows workers to resign in the midst of a strike and scab on coworkers. A huge number of professionals and supervisors were even deemed exempt from representation (Lichtenstein 66). Legislation and corporate wealth are eroding the power of organized labor and thereby obfuscating workplace democracy; extinguishing employee rights; eroding the living standards of working, working- poor, and middle class Americans; muting the voice of minorities; retarding environmental improvements; increasing corporate domination of politics; and auguring exploitation of workers throughout the world. However, a significant portion of freedoms, to which Americans have become accustomed, would be greatly diminished or non-existent without the social values that are embodied by organized labor. Evidence suggests that employers seldom behave democratically without the mandate of a higher authority such as the government or a union. It is no wonder that workplace dictatorships are becoming a widespread phenomenon as government regulations fail to adequately protect workers but enhance the power of employers: â€Å"Fear of being fired, downsized, laid off, of not making pension time, poverty in a new economy, of part-time and insecure, low-paid jobs, an... ...titute, 1997. Lawrence, Vince. â€Å"John Sweeney’s Militant Unionism.† The New Republic 6 Oct. 1997: 23 - 24. LeRoy, Greg. â€Å"The Terrible Ten.† The Progressive 28 May 1999: 27 - 30. Lichtenstein, Nelson. â€Å"Work Rights, Individual Rights.† Dissent Spring 1997: 66 - 72. Mantois, Gregory. A New Labor Movement for the New Century. New York: Monthly Review Press, 1998. Moberg, David. â€Å"Union Pension Power.† The Nation 1 June 1998: 16 - 19. Puddington, Arch. â€Å"Is Labor Back?† Commentary July 1998: 39 - 42. Shribman, David. â€Å"Big Labor Gets Its Act Together.† Fortune 29 Sept. 1997: 60 - 61. Silbiger, Stephen. â€Å"State of Unions.† National Review 26 Jan. 1998: 20 - 21. Smith, Peter. â€Å"The Fractured World of the Temporary Worker.† Journal of Labour 22.2 (‘1998): 414 -427. Wells, Don. â€Å"Labour Solidarity Goes Global.† Canadian Dimension 32.2 (1998): 33 - 39.

Business Structures Essay

If someone wants to start a business, that person would have to decide which structure he or she would want to use. To know what kind of structure he or she has to know what kind of business he or she is trying to run and who will run it with him or her. Structures range from sole proprietorships and partnerships to corporations. When companies first start up, they consider sole proprietorships or partnerships, but as they grow into larger companies, they become corporations. When someone first opens a business it may just be him or her, those companies consider sole proprietorships, meaning a business owned by one person. There are several advantages when running a sole proprietorship. First is it is the simplest type of business to start and run and also it is not regulated as much. Also, sole proprietorships pay lower income taxes compared to other business structures. Last, when a company is a sole proprietorship, decision-making rest on the owner’s shoulders but the owner keeps all profits (Films Media Group, 2011). Even though there are some very good advantages owning a sole proprietorship, there are also some disadvantages. The biggest advantage is also the biggest disadvantage, the bills, debts, and major obligations. If the owner does not pay the bills, the company cannot run. Another big disadvantage is the capital the business has. If the owner does not have some other capital invested into the company, the company will not grow. The last disadvantage is running the company when the owner leaves or dies. Because there is no one else with stock, the owner has to let the company go (Parrino, Kidwell, & Bates, 2012). The next business structure is a partnership, which consist of two or more owners that legally run the business together. With a partnership, the owners know what his or her position in the company is and how the profits would be split. There are two different partnerships; a general partnership and a limited partnership. The general partnership has the same advantages as a sole proprietorship, but has one more disadvantage. All owners have unlimited liability regardless of  the percenta ge in the company (Parrino, Kidwell, & Bates, 2012). To avoid the big issue with a general partner, the partner would sign into a limited partnership. In a limited partnership, the company can have general and limited partners. One or more general partner has unlimited liability while limited partners only deal with the obligations he or she provides. To qualify as a limited partner, he or she cannot engage in the managing of the business (Parrino, Kidwell, & Bates, 2012). The last business structure is the biggest and the last to get to. Large companies consider themselves to be corporations. Corporations consider themselves as a â€Å"person,† where the corporation can sue and also be sued, borrow money, and own assets like real estate. An advantage of a corporation is that the stockholders have a limited liability for all of the corporation’s obligations. Because the corporation is a â€Å"person,† the corporation is taxed as a â€Å"person† on the income it earns (Films Media Group, 2011). References Films Media Group (2011). Planning Your Business: Research, goals, and business plans [Video podcast]. Retrieved from https://newclassroom3.phoenix.edu/Classroom/ToolContainer.jsp?context=co&contextId=OSIRIS:40817068&activityId=16e92012-daa3-4692-89b0-622c50a227b6&profileId=4136b5d5-519c-4e35-a63a-f84741e11cd2&syllabusId=OSIRIS:40817068&version= Parrino, R., Kidwell, D. S., & Bates, T. W. (2012). Fundamentals of Corporate Finance (2nd ed.). Danvers, MA: John Wiley & Sons, Inc..

The Complications Of Success By Russell Banks - 1647 Words

The Complications of Success Most of the stories in the book Success Stories by Russell Banks seem to be about failures. The stories are about failures because the author shows that failures can lead to success based on what you do after failing, the reasons some may fail at becoming successful, and how certain ideas of success that other people have for you, can pull you away from your own ideas of success. Failures can either motivate you to try harder for success, or you can choose to let failures that take place make you unsuccessful. In these story’s it seems that the main character cannot become successful, because he has failed repeatedly at†¦show more content†¦Being in poverty, and not having parents who are successful, may be why Earl cannot become successful. Simply because while growing up success seemed so far out of reach. This could give Earl the idea that success just isn’t possible for him, because it didn’t happen to the people who rais ed him. It didn’t happen to the people who raised him because his father let alcoholism prevent him from being successful, and it didn’t happen to his mother because of the situation she had been put in at a young age, with kids, a horrible husband, and surely other reasons that can come along with those things. Although earl does not have the same problems that his parents did or do when it comes to things that prevent them from becoming successful, he lets their struggles of becoming successful prevent himself from overcoming his struggles of becoming successful. Earl lets his lack of perseverance, and inability to realize his potential get in the way of him being successful. If earl could see the potential in himself that others see in him, and realize that he needs to not just give up on his aspirations and continue to strive, he would use that potential and perseverance in the right ways and become successful. 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