A Quirky Inspiration free essay sample

To inspire a person is a most powerful action. Through inspiration one can realize his or her dreams and life-long goals. For most people a family member, such as a mother, father, big brother/sister, or war-veteran grandfather would be their inspiration. As for me, even though my mother and father were both great influences, my greatest influence came from a television program. Nearly everyday after school, I would come home, snuggle into the comfort and warmth of the couch, and turn on the television. Upon hearing the fast paced dialogue, seeing familiar faces, and hearing the theme song, a sense of relief and total tranquility would wash over me. All the day’s hardships would disappear. I would forget about any former stress, and I would put off making calls to my friends to chat about all the gossip I had heard at school earlier that day. For when five o clock rolled around, I was planted in front of the TV, only to watch another gratifying episode of Gilmore Girls. We will write a custom essay sample on A Quirky Inspiration or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page Gilmore Girls is what I had to look forward to in my day, if not because I was having a bad day, then because I simply loved to laugh at the quirky remarks and the sarcasm etched in each characters face and spoken in their dialogue. Each of the characters was unlike anyone I had ever known. Each character had an especially unique quality that made him or her fit into the town of Stars Hollow (often referred to as an â€Å"asylum†). The more I tuned in, the more I found myself wanting to be a Rory Gilmore: she was smart and funny; she had the kind of relationship with her mother that many girls would kill to have. The show brought a new light into television: it focused on the shy, sensitive, book-smart girl, instead of the misunderstood, beautiful, popular girl. I quickly found a character that I could identify with. This convinced me that I wanted to be exactly like her. Much of the credit for Rory’s success came from her mother, who fed her classic novels, along with meals at Luke’s Diner. In the process of getting into Harvard or to any college in fact, I recently have discovered is a long and grueling process. My mother has always pursued me to follow my dreams and to make my own decisions. I am a particularly indecisive person, so undertaking this task was not easy for me. In growing up and developing into her own person, discovering what she wanted to be and how she was willing to achieve those dreams; that was how Rory Gilmore inspired me. Being known as the ‘book-smart girl’ while all the while maintaining her friendship with Lane, going on outings with her mother, dating, and managing to make it to Friday night dinners with her grandparents has shown me that education and personal lives can come together. Although I am only one girl who is still trying to find her place, Rory made me feel as if I could conquer anyt hing, from studying for mid-terms to obtaining the college of my dreams. Disregarding which college I may choose, I will always know that the road, to which I traveled to get there, was a road well worth traveling.

Free Essays on George Kennans American Diplomacy

Critiquing American Diplomacy George Kennan’s American Diplomacy provides anyone studying American politics with an accurate glimpse of American ideology, misperception and thought in the first half of the twentieth century. Even though this book is a collection of lectures where there is obvious subjectivity, Kennan successfully displays an objective view on the topics he discusses in the lecture. Kennan’s lecture gives the reader a panoramic view and solid background of American thinking about themselves and the rest of the world, with respect to events leading up to the conflict with the Soviets. As a result, Kennan gives a clearly articulated picture of his thoughts. Kennan explains his ideas in a manner which the average college level student could understand, but does it in a sophisticated manner, which is refreshing. Kennan compares American security ideas to the Roman Empire because Americans had a sense of security and superiority that had not been seen in the world since those times. He compares American ideas and situation in 1950 versus American ideas and situation in 1900 and discusses this from a subjective and objective point of view in order to back up his opinion and present a clear case without discussing the situation at a complete slant, which I think shows how qualified he was to influence American foreign policy because he had every right to speak about things subjectively because he was such an authority and had good insight due to his ability to look at the situation from every perspective, not just the American perspective. I believe Kennan attempting to be as unbiased as possible in this lecture allows him to give anyone reading the book a good chance to develop their own argument, which is a good quality because he is not trying to force his opinion on anyone. â€Å"†¦our national consciousness is dominated at present by a sense of insecurity greater even than that of many people of Western E... Free Essays on George Kennan's American Diplomacy Free Essays on George Kennan's American Diplomacy Critiquing American Diplomacy George Kennan’s American Diplomacy provides anyone studying American politics with an accurate glimpse of American ideology, misperception and thought in the first half of the twentieth century. Even though this book is a collection of lectures where there is obvious subjectivity, Kennan successfully displays an objective view on the topics he discusses in the lecture. Kennan’s lecture gives the reader a panoramic view and solid background of American thinking about themselves and the rest of the world, with respect to events leading up to the conflict with the Soviets. As a result, Kennan gives a clearly articulated picture of his thoughts. Kennan explains his ideas in a manner which the average college level student could understand, but does it in a sophisticated manner, which is refreshing. Kennan compares American security ideas to the Roman Empire because Americans had a sense of security and superiority that had not been seen in the world since those times. He compares American ideas and situation in 1950 versus American ideas and situation in 1900 and discusses this from a subjective and objective point of view in order to back up his opinion and present a clear case without discussing the situation at a complete slant, which I think shows how qualified he was to influence American foreign policy because he had every right to speak about things subjectively because he was such an authority and had good insight due to his ability to look at the situation from every perspective, not just the American perspective. I believe Kennan attempting to be as unbiased as possible in this lecture allows him to give anyone reading the book a good chance to develop their own argument, which is a good quality because he is not trying to force his opinion on anyone. â€Å"†¦our national consciousness is dominated at present by a sense of insecurity greater even than that of many people of Western E...

A 2,000 report exploring and critically analysing the complex needs of Essay

A 2,000 report exploring and critically analysing the complex needs of offenders and rehabilitative practices which promote desistance - Essay Example These may include substance abuse problems, unemployment or financial problems, difficult family backgrounds, homelessness, or even psychiatrist problems. It is important for the institutions management to acknowledge that these factors are interlinked in most cases and will vary mostly along age and gender lines (David 2004; Warr 2002). Theoretical evidence affirms that a combination of individual and social factors can be linked to an increased probability of reoffending and should be routinely reassessed. Also referred to as ‘criminogenic’ needs, they can be attributed to certain types of crime. For example, heroin use is closely linked with shoplifting and other acquisitive offending while alcohol binge drinking is often associated with violence and other petty crimes. These factors can be further classified into static or dynamic. Factors such as employment, drug misuse, or education are subject to change and are thus, classified under dynamic while strong predictor s of reoffending such as gender, age, or criminal history are static. So, what helps individuals stay away from crime? Maruna et al (2008) propose that desistance can only work out if the complex needs of the offenders are met. As such, there is a strong correlation between the social or external aspects of an offender’s life (such as the support of those around them), as well as, the psychological or internal (what they subscribe to and what they want to achieve with their lives) and relapsing or desisting. Factors that might work in favour of desistance are such as: i) Family and relationships: studies have suggested that establishing supportive bonds with spouse or family members appears to boost chances of desisting from crime on condition that the family members are not involved in crime or substance abuse themselves. These good relationships are crucial to recidivism owing to a number of reasons. First, they minimise the amount of time and interactions spent with

Kidnapping tourists in mexico Essay Example | Topics and Well Written Essays - 1750 words

Kidnapping tourists in mexico - Essay Example The article also has an overview of some of the most dangerous countries for tourists to visit, such as North America/ Mexico, South America, Africa and Asia. Analysis – The article was quite useful and could be used for supplying a lot of information on the subject. Reflection – I found the article interesting and understood that the author has taken a lot of trouble gathering important information. 2. Iuri Tarabanov (2008) World’s Most Dangerous Countries for Tourists. Travel Web Directory. Summary This is a web article that reports on Travel news. The author Iuri Tarabanov sheds light on the world’s top 10 most dangerous countries for tourists. The data collected is from â€Å"iJet Intelligent Risk Systems† and the renowned magazine â€Å"Forbes† has contributed by composing the level or rating of the most dangerous countries for tourists. The agency took into account factors such as security services, terrorism, kidnapping, civil disorder s and other criminal and geopolitical stability factors while doing the rating. Some of the most dangerous countries in the world as noted by the author were Somali, Iraq, Afghanistan, Haiti, Pakistan, Sudan, The democratic Republic of Congo, Lebanon, Zimbabwe and Palestine. Analysis The article specifies the different types of dangers significant to each of the above countries and sheds light on the type of danger that is prevalent in these countries thereby creating an awareness on the dangers that tourists are exposed to if they visit them. Reflection The data that was given by the â€Å"iJet Intelligent Risk Systems and â€Å"Control Risks† agencies was not only enlightening but it also served to kindle awareness among the people of the different kinds of risks involved while visiting different countries. This type of article would surely be very useful in my study on dangerous countries for tourists. 3. The Washington Post Newspaper, Athens Travel Forecast. Good Deals f or Tourists who don’t mind occasional risks. Summary The Washington post news article sheds light on the tourism in Athens, Greece. It talks about the beautiful archipelago, beaches and buildings which is a welcome escape for tourists to relax and enjoy themselves. At same time, it warns tourists of impending dangers in the form of strikes and violent protests that suddenly erupt and causes major problems for tourists. The article in the Washington Post also discusses the best restaurants for food, but at the same time it creates awareness on the rising crime in the city and the drug trafficking that takes place there. Analysis The article in the Washington Post gives more details on the beauty spots, food and restaurants found in Athens Greece, but however shed some light on the dangers that the tourists may be exposed to. Though one cannot say that Greece is a highly dangerous country, yet the paper creates an awareness about what to expect if one visits the place. Reflecti on Though the Washington Post is a reliable newspaper, this particular article is not very relevant to my research study because it more describes the beauty and hotspots of Greece but has little on the dangers tourists might be exposed to. However, the article would be partially useful to include in my study. 4. John Keay (2010) The Mammoth Book of Travel in Dangerous Places,

Formal report Assignment Example | Topics and Well Written Essays - 2000 words - 1

Formal report - Assignment Example This paper will give an overview of what social networking is about, what it is used for and will endeavor to give examples of companies and their social networking strategies. A social networking plan and recommendations will also be given specifically for Stone Shine, a stone cleaner product that the company, Deb Richey Co. launched online. A social network service is defined as â€Å"the grouping of individuals into specific groups, like small rural communities or a neighborhood subdivision† (Social, 2011). While the networking can be done physically such as in a workplace, or at school, social networking is most popular online. This is because the internet has transcended physical barriers and has made it possible for millions of people from all over the world to come together for a shared interest. Connecting is no longer impeded by the inability to physically come together. Social networking websites function like an online community of internet users (Social, 2011). Members communicate by email, forum posts, blogs, comments and instant messages. These are all features that can be made available by the varied social network sites. By becoming a member, these sites give every individual the chance to create his/her own profile. Having a personal profile lets members put personal information and define their personal interests, this is what sets social networking platforms apart from offline ones. There is ease of access to member information that makes it accessible to other members, making it easier for people to interact. The networking part also does not have to happen in real-time. This takes away the pressure of having to interact within a time frame, as with personal meetings, or whether to even react at all. Because there is this absence of pressure to connect plus the ease of access, social networking is a very enjoyable activity for lots of peo ple. Coupled with the fact that man is a social

Expression and Purification of recombinant Green Fluorescent Protein

Expression and Purification of recombinant Green Fluorescent Protein ABSTRACT: The purpose of this experiment was to determine if a His-6 tagged recombinant form of Green Fluorescent Protein could be expressed in a pRSETA vector of E. Coli. This was determined through multiple procedures beginning with purifying the sample with Ni +2 agarose chromatography which showcased the relative fluorescent activity of the samples, which elution sample two (E2) had approximately 100,592.2 RFU/mg . The yield of total protein was found by use of a Bradford Assay and a standard curve. The purity of the GFP was determined by comparing the intensity of bands that appeared at around 31.4 kDa (the molecular weight of rGFP) to a molecular weight ladder on an SDS-PAGE gel. The Western Blot test, utilizing a nitrocellulous membrane, confirmed the expression of rGFP. The Western Blot confirmed that the correct bands were analyzed in the SDS-PAGE gel which E3 had an estimated purity of 0.4, indicating a yield of approximately 29.4 ug of rGFP for the third Elution (E3) after a total p rotein amount of 49 ug was extrapolated from the Bradford assay. INTRODUCTION: The Green Fluorescent Protein was first isolated from the Aequorea Victoria jellyfish and studied by Osamu Shimomura. In other organisms other than this specific jellyfish, there are fluorescent proteins with more color varieties than just green (Ward). GFP is able to fluoresce due to the formation of a chromophore in its center that resides in a stable beta barrel structure. The interactions between Ser64 and Gly67 on an alpha strand form a five member ring structure and its this structure that contains properties for fluorescence. When ultraviolet light is absorbed at 395nm, an emission wavelength of 510nm is seen as fluorescent green light. The excitation wavelength of 395nm excites the electrons within the protein and raises the energy of the protein. As the protein loses energy, it emits the energy at a wavelength of 510nm. Wild type GFP does not have immensely visible fluorescence, therefore a genetically modified form of GFP, GFPuv, was used in order to better observe fluoresc ent activity. For rGFP to be fully expressed, GFPuvs open reading frame was pRSETA vector, which was cut by a restriction enzyme at sites that complimented the start and stop codons of the His-6 tag sequence. A T7 promoter and Xpress Epitope were also genetically added into the vector with the purpose of simplifying purification. T7 polymerase minds to the T7 promoter and activates the production of GFP, while the Xpress Epitope allows for the binding of a primary antibody. The binding of a primary antibody allows for the binding of a secondary antibody which contains horseradish peroxide, producing bands on the nitrocellulose membrane. (2 pg 351) Histidine-6 tags primary purpose is to aid in the purification of rGFP proteins in affinity chromatography. Histidine has an affinity to bind with transition metal ions, and is why a Nickel+2 agarose chromatography was chosen. When a sample is passed through the Nickel+2 agarose column, the His-6 tags fused with rGFP protein will bind to the column and any proteins not of interest or containments will pass through the column. To elute the bound rGFP proteins from the column, elution buffer containing Imidazole is used and competes with the histidine residues for the binding sties to the Nickel+2 agarose column due to its higher affinity for them. Elutions from the column therefore contain rGFP. (2 pg 359) The purpose of this experiment was to see if a His-6 tagged recombinant form of GFP, from the E. coli strain BL21pLysS, could be expressed and then purified using Ni+2 agarose affinity chromatography technology. If this experiment is successful, an inexpensive and relatively simple method to measure gene expression and locate and track proteins may be at hand. MATERIALS AND METHODS: Bacterial Expression Grow two bacterial cultures: G, containing BL21, DE3, pLysS, and the plasmid pRSETA-GFPuv with the GFP sequence, and V, which is identical to G, but it does not have the GFP sequence. Incubate both cultures at about 37Â °C until OD600 equals 0.5. Transfer 1ml of each culture into separate centrifuge tubes and centrifuge to obtain a pellet. The supernatant from the tubes will be discarded and then label the tubes V0 and G0 and stored at -20Â °C. Induce the rest of the culture with IPTG and allow to grow for 3 hours. After the 3 hours, collect 1ml of each culture and centrifuge. Label the tubes V3 and G3 and were store at -20Â °C. Collect 15 ml of G and centrifuge, and label it as G3-15ml and store at -20Â °C. (1 pg 105) Preparation of Crude Extract After the slow freeze process, add 1 ml of breaking buffer (10mM Tris, pH 8.0; 150mM NaCl) to the frozen bacterial pellet G3-15ml, and pipette up and down until completely dissolved. Then transfer the contents to a centrifuge tube and vortex for 5 minutes and place in a 37Â °C water bath for 10 minutes. Afterwards, incubate in dry air at 37Â °C. Centrifuge the mixture at 4Â °C and transfer the supernatant into a clean centrifuge tube and take a small sample to label as GCE. The remaining supernatant will go through the Nickel+2 agarose column. (1 pg 110) Ni+2 NTA Chromatography Pack a 3ml plastic syringe with a small amount of glass wool to prepare the Ni+2 agarose column. After that, pipet 1 ml of breaking buffer into the syringe to remove any air bubbles. As the breaking buffer is going through the syringe, pour some breaking buffer into the leur-lock and then screw it onto the syringe. Add 1ml of 50% Ni+2 agarose into the column and then open the leur-lock to pack the agarose matrix in the colum. Add 5 ml of breaking buffer subsequently into the column to push the ethanol of the Ni+2 agarose through the column. Then close the leur-lock and apply the crude extract to the column and allow 5-10 minutes for the histidine tags on the rGFP to bind with the Ni+2 agarose beads. Open the leur-lock and collect the non-crude extract in a centrifuge tube labeled W1 for the first wash. Add 0.5ml aliquots of breaking buffer to the column and collect the washes in tubes W2-W10. Add 10 increments of 0.5ml of elution buffer and collect in tubes E1-E10. Then store your GC E, W1-10, and E1-10 samples at -20Â °C. (1 pg 111-2) Bradford Assay Analysis To create a Bradford assay, mix protein sample with water, and then add Bradford reagent dye. Vortex the mixture and then transfer it to the wells of a microtiter dish, so that the absorbance of your protein sample can be determined at 595nm using a spectrophotometer. To determine the amount of total protein present in the sample volume, you must create a Bradford standard curve using 0, 2.5, 5, 10, 15, and 20 ug of BSA and the absorbance values observed to determine a best-fit line. Perform the assay in singlicate using samples W1-W6 and E1-E6 to determine if the microplate data falls within the standard curve. Once you have determined what volume of sample to use, repeat the Bradford assays two more times for each sample. Use the standard curve and experimental absorbance values to extrapolate the total protein present in the volume of sample you use. (1 pg 124-6) SDS-PAGE/Coomassie Blue Analysis of rGFP Fractions Create a 12% Resolving Gel by mixing water, 30% Acrylamide, 10% APS, TEMED, and 4x resolving buffer [0.75M Tris pH8.8, 0.4%SDS]. Pour the resolving gel into a gel electrophoresis set up and overlay with some water. Allow the resolving gel to polymerize and proceed to making a 5% Stacking gel by mixing water, 30% Acrylamide, 10% APS, TEMED, and 4x stacking buffer. Pour the stacking gel on top of the resolving gel and immediately insert a comb to form the wells. Prepare your loading samples with 4x sample loading buffer and then vortex, boil, and centrifuge. Do this for the G0, G3, GCE, W2, W3, E2, and E3 samples (or fractions that contained the most rGFP fluorescence). (1pg 111-2) After the gel has solidified, transfer the apparatus into the electrophoresis tank and load the electrophoresis buffer into the tank. Then load your samples into lanes one through seven respectively along with a marking ladder to compare the distances the samples traveled after electrophoresis. Electrophorese at 200V for approximately 45 minutes and then stain your gel with Coomassie Blue dye. To remove the stain that has not been absorbed by proteins, the gel is soaked in dilute acid and methanol. Western Blot Develop an SDS-PAGE gel as previously described using your sample fractions and transfer the proteins onto nitrocellulose transfer membrane using two locking cassette lids with sponges and filter paper encompassing the gel and nitrocellulose. Remove the nitrocellulose and stain the membrane with Ponceau S stain, and allow to incubate for approximately 2 minutes. Once stained, wash the membrane with nanopure water until bands are visible on the membrane. Next, perform the blocking step by placing the membrane in a container containing 5% non-fat dry milk/TBS solution and incubate on a shaking platform. Pour out the blocking solution and add 0.05% Tween 20/TBS solution and then allow it to incubate on a shaking platform, and then pour out the solution. Repeat this wash step two more times. Add mouse IgG anti-Xpress epitope MAb and allow incubate on a shaking platform this is the primary probe step. Then repeat the wash step as mentioned previously three more times, and add Sheep IgG a nti-mouse IgG conjugated horse radish peroxidase polyclonal anti-serum solution (the secondary probe). Allow the membrane to incubate, and then perform the wash step twice. For the final wash, use only TBS, and then add TMB substrate solution and incubate until appropriate banding is shown. Stop the reaction with distilled water and scan or photograph the membrane as the results may fade with time. RESULTS: In the specific strain of E. coli, BL21pLysS, pRSET-GFP represses the lac promoter which is part of the operon needed for T7 polymerase activity. T7 polymerase binds to the T7 promotes of pRSET-GFP and activates the His-6 tag that expresses the fluorescence of GFP. Even though some GFP is produced this way, it is experimentally not enough, so IPTG is added to repress the repressor so an increase of GFP production can occur. V0, V3, G0, and G3 were all bacterial cultures of E. coli with specific distinctions between all of them. V0 and G0 were collected from bacterial cultures that were not yet induced, but G0 contained the GFP sequence while V0 did not. V3 and G3 were the V0 and G0 samples after induction had occurred for 3 hours, and again G0 contained the GFP sequence while V0 did not. W1-W10 were washes collected from the Ni+2 agarose column using the breaking buffer whereas E1-E10 were washes collected using elution buffer. Figure 2. SDS-PAGE/Coomassie Blue stained gel analysis of rGFP SDS-PAGE gel, which was used to analyze our purification of rGFP from a crude extract, and the corresponding molecular weight. This gel was made with a 12% resolving gel and a 5% stacking gel. Wildtype rGFP has a molecular weight for 27kDa. The ladder has kDa rungs of 97.4, 66.2, 45.0, 31, 21.5, and 14.4. The band for rGFP has been indicated on the figure. The different samples were taken during our purification. The highest protein containing washes and elutions were used in the SDS-PAGE gel. G0 represents a sample containing rGFP prior to induction. G3 represents a sample containing rGFP after 3 hours of induction. GCE represents the GFP crude extract after the slow-freeze/quick-thaw process. Figure 3. Western Blot of rGFP We used the Western Blot to detect, and more importantly, quantify proteins that react with antibodies. The Ponceau S stain was performed to highlight the existence of our proteins successfully being transferred from the SDS-PAGE gel. We then performed a blocking step with 5% non-fat dry milk/TBS solution followed by washes with 0.05% Tween20/TBS. The probe first used was mouse IgG anti-Xpress epitope MAb solution, which we followed with washes of Tween20/TBS solution. The second probe used was sheep IgG anti-mouse IgG conjugated horseradish peroxidase polyclonal anti-serum solution, which we followed with two washes of Tween 20/TBS solution with the third wash being pure TBS. Then, TMB solution was added to develop the nitrocellulose membrane and this reaction was stopped by pouring distilled water over the membrane. Above are the 8 lanes, G0, G3, GCE, W2, W3, E3, E4, and a known molecular weight ladder, respectively. CONCLUSION/DISCUSSION: A great amount of research has been put into the field of monitoring gene expression and there are currently many research laboratories that use labeled antibodies and other various means to do so. However, this new recombination technology, a vector can be created to provide a simpler and less expensive approach that can even be performed in vivo. The purpose of this experiment was fulfilled as His-6 tagged rGFP from a particular E. coli strain was successfully expressed, purified, qualified, and quantified by means of Ni+2 agarose chromatography. GFP was initially expressed as a histidine tagged protein that was inserted in E. coli and subsequently a crude extract of this protein was isolated. Throughout the experiment, we were able to inspect and measure the fluorescing activity of our elutions with the use of handheld UV lights. GFP was successfully expressed because of the fluorescent activity that was emitted when shown under UV light. This rGFP was purified using a Ni+2 agarose column which allowed for the binding of the His-6 tagged rGFP to the Ni+2 agarose beads in the column, and subsequently let all other proteins not of interest and containments to be collected in washes. Then, rGFP proteins were eluted by passing elution buffer containing Imidazole through the column because Imidazole has a higher affinity for the Ni+2 agarose beads than does Histidine. Therefore, one could expect that most if not all of the rGFP would be found in the elution washes and would present higher fluorescing activity than the washes with breaking buffer. Figure 1 clearly supports this argument. From referencing Figure 1, the most rGFP was found in E2 because it exhibited the highest fluorescent activity in RFUs. Using a standard curve based off of known quantities of mass of a certain protein, one is able to extrapolate the amount of protein present in ones sample. This can be used to estimate the amount of rGFP activity. For instance, E2s specific activity was found to be approximately 100,592.2 RFU/mg. W1-W6 had the least amount of fluorescing activity because the washes were done prior to the addition of Imidazole through the column. This is expected as Imidazole would flush out rGFP in the elutions, which would produce the highest GFP fluorescing activity. Percent purity of the rGFP samples were approximated using the SDS-PAGE gel with Coomassie blue staining to allow visible bands to appear which were used for comparisons. The molecular mass from each individual sample was determined by comparing the bands with the bands of the known weight ladder. The percent purity of each individual band was determined by comparing the bands against each other. The molecular weight of wild type rGFP is 27 kDa, and the molecular weight of rGFP for mutant rGFP used for this experiment was approximately 31.4 kDa with the 40 extra amino acids that were added taken into account. For example, E3 percentage of purity was found to be .4. The amount of protein yielded was 29.4 ug of rGFP for the third Elution fraction (E3) after a total protein amount of 49 ug The Western Blotting technique detected and quantified proteins that reacted with a specific antibody. In our case, it verified rGFP was expressed and whether or not it fell in the range of the expected molecular weight of 31.51 kDa. Lanes GCE, G0, G3, and E3 all contained a bright band around 31.4 kDa, indicating that rGFP was existent. This result corresponds to the expected of E3 having the one of the highest rGFP activity. The fact I had used E3 for the SDS-PAGE and Western Blotting skewed my results, but E2 and E3 had the highest activity for me. Even with my error, GFP was effectively expressed and purified and its finding is important to science because it can be used to track cancer causing cells one day to help figure out where problem spots begin and can help eliminate the issue before it escalates indicating that fluorescent proteins might eventually be clinically useful in cancer patients (Hoffman). This can occur since GFP can successfully be expressed in other organisms as this experiment through the use of E.coli, humans cells can certainly be tagged and label the tumour before treatment and then monitor for fluorescence after treatment to identify possible recurrence or metastases (Hoffman). The same can be said with heart disease and other health related problems. The future of GFP remains limitless when scientists can unlock its full medical potential. REFERENCE/BIBLIOGRAPHY (1) Rippel, Scott. BIOL 3380 Fall 2010: Biochemistry Lab Manual. The University of Texas at Dallas. Richardson, TX. (2) Rippel, Scott. BIOL 3380 Fall 2010: Biochemistry Lab Lecture Notes. The University of Texas at Dallas. Richardson, TX. (3) Ward, William. History of GFP and GFP Antibodies. Brighter Ideas. 2009. 26 Oct, 2010. http://www.brighterideasinc.com/proteins-antibodies/history-of-gfp-and-gfp-antibodies/ (4) Ninfa, Alexander J. and David P. Ballou (1998). Fundamental Laboratory Approaches for Biochemistry and Biotechnology. (5) Hoffman, Robert M. Uses of Fluorescent Proteins to Visualize Cancer In Vivo: Can Fluorescent Proteins be Used in Humans? 2005. 28 Oct. 2010. http://www.medscape.com/viewarticle/513975_6

Women’s New Role Essay -- Essays Papers

Women’s New Role In coming to understand what it meant to be a girl I was affected by my era, women’s sports, and the place that I was raised, as much as the independent feminist spirit that my mom was exposed to and possessed. My mother’s mom raised her to believe and accept the traditional female role. My grandmother put emphasis on the women staying home and existing solely as a support system for the man. She found comfort and security knowing that it was the man’s burden to â€Å"bring home the bacon.† My mother, however; was influenced by the time and took it upon herself to take care of herself and make her own money. At a young age she along with many other women of her time wanted to rebel against the expected role and thus have a more fulfilling life. Family and children were an important priority for her but she felt strongly that she could do more. Her parents had a wonderful and loving relationship but her father had three massive heart attacks at age 42 a nd was not suppose to live a year. My mother was one of the oldest of eight children and was well aware of her mother’s very frightening predicament. This aforementioned life experience and her inherent desire to educate herself made her believe that women could do more. My mother did want to marry but wanted to be in the marriage because she wanted to not because she had to stay in an unpleasant situation. She wanted to make it on her own. She did not want a man to control her and most of all to tell her what to do. Her competitive drive for success in the business world was luckily passed on to me and gave me the confidence and perseverance that I need to get through life. My mother taught me that I was a woman, so things may be harder and that I must never e... ...s influential as my role models on the women’s US national team. They all gave me the courage and confidence in myself as a woman. Today I am happy to see that women are coming together and trying to inspire each other to take on the world. By simply typing in â€Å"Women Role model† in goggle I found several sites for women engineers, doctors, and writers trying to make something out of their lives. I was happy to see that someday when my daughter is trying to pursue her dream that she will have options because of the risks these women took. And I will be proud to be able to say that I was one of those women. Work Cited Dorance, Anson. Telephone interview. 29 May 2003. Gonzales, Monica. Personal interview. 30 May 2003. Hamm, Mia. Go for the Goal. HaperCollings. NY 1999. Lilly, Kristine. Personal interview. 30 May 2003. UNCtarheel.com.May 31 2003.

Dehumanization in Slavery based on Accounts from Clotel

William Wells Brown warns us that slavery does not only victimize the weak and blacks. In his novel, he points out that â€Å"no one is safe† from slavery, whether â€Å"white or black† (Schweninger 23). In the face of this fact, it seem ironic that the country that is now called the â€Å"land of the free† and â€Å"home of the brave† was once a country where slaves were sold and treated almost with cold blood (Schweninger 23). Indeed, no one is safe, for even a president’s daughter has been sold to be a slave. What could be more painful than to be parted from your beloved and your daughter and be sold as a slave, knowing that your daughter would be treated as ill as a servant? This is the Calvary that Clotel had to bear as a slave, though she was, by popular lore, a daughter of the president. And if it were true that she was indeed a daughter of the president (Schweninger 28), would it not be more painful on her part, knowing that she was the daughter of the most powerful man in country, yet she had to be a slave and a concubine? The very thought of this situation dehumanizes Gods masterpiece. There is a rich literature about Brown’s novel, both digital and printed. Most of them bank on the cruelty of slavery. Articlemyriad. com reminds the readers that the novel is fiction, but description of slavery with all its vividness in the accounts of the characters’ lives is true. It can be well understood how something fictitious can capture the truth behind every word and expression. The writer himself was once a slave, and in fact, the first part of the novel is a biographical account of how he escaped slavery (Analysis and Themes 1). Hence, we see slavery, not in the eyes of someone who has just seen it, but someone who had been a slave, someone who had as firsthand experience of the dehumanizing treatment to slaves. It is not, however, the hard work that Brown resents, but the fact that humans, free or slave are not to be sold (Schweninger 22). At this juncture, it is important to focus more on the historicity of the accounts in the novel. As Articlemyriad. com points out, the novel is fiction, but the accounts are real. No president of the United States would have had a slave daughter, but the experiences of the characters in the novel, from the most â€Å"optimistic† to the most â€Å"pessimistic† were not products of imagination (Analysis and Themes 3). As Clotel became a mistress, and was sold, later tried to escape and then commit suicide in despair, Brown was showing a concrete example of what happened to slaves. It was not their fault to be born as mulatto or black or yellow skin, but they faced the discrimination and social prejudice and stereotyping. One example of this stereotyping is stated in the website mentioned. Society does not frown upon the man who sits with his mulatto child upon his knee whilst its mother stands a slave behind his chair† (Brown 55). We can see from this statement that there is something bout being a mulatto child and a slave mother. A slave, though mother of the master’s child, remains to be a slave and does not seem to have the right to be wife, and only has the right to be a mistress, a whore. If a white were mother to child of a master, then the master would have to marry her in most cases, of the master would have to face suit, but slaves had no liberty to fight for their rights no matter how disgraced they have been. Though the article points out that the novel is fiction, it gives way to Brown’s claims that the novel is â€Å"no fiction† (Analysis and Themes 2), for it is â€Å"founded in truth† (Analysis and Themes 2). The truthfulness of accounts in the novel is what the other two articles by Schweninger and Giulla, although Giulla’s accounts are rather more focused on the language of the blacks. Schweninger puts forth that in as much as the events in the novel could have taken place to slaves; the accounts in the novel were not historically accurate as far as chronology of events is concerned. She mentions several inconsistencies in the chronology of events, one of which is the impossibility of Clotel’s birth in 1798 and â€Å"have her mother witness the 1842 burning when her daughter was only 16† (Schweninger 27). Therefore, according to Schweninger, the point of Brown is not to make a accurate historical account (although he told the readers that the novel is not fiction), but to stress the inhumane treatment to slaves, especially to African Americans and the mulatto. In fact, in the preface of the novel, Brown states â€Å"Were it not for persons in high places owning slaves†¦ Slavery would have long been abolished. (Schweninger iv). And who were these persons in high places? Brown identifies them as â€Å"professed Christians† who give the system (slavery) â€Å"a reputation† (Schweninger iv). Schweninger suggests that the â€Å"undercutting of the chronology of events stresses that it is not the chronology that is important in the no vel but the universality of the topic and slavery in any year or era would have been the same† (Schweninger iv). The people would have suffered just as much and the abusers would have abused the slaves just as much. And as Brown has said, slavery remained not condemned, because the people who had the power to stop it knew its evil but did not want to lose their slaves. They would be hurt in material loses. The battle cry of Brown is not for people to be freed from work. Everyone has to work, but no one deserves to be sold as a slave. Schweninger traces the story of Clotel to another popular lore called Child, where Clotel was originally named Rosalie. She attests that the story of Child was copied word for word except for some insertions by the Brown to express his political views (Schweninger 28). For instance, Scweninger narrates, when Clotel urged the master to move from one place to another, Brown inserted some statements that would highlight the anti-slavery theme of the novel. So, in the original text from Child, the line says â€Å"the slave mistress urged her master ‘to remove to France or England (Scweninger 64)†, but in the text of Brown, there is the insertion â€Å"where both her child would be free and where colour was not a crime (Brown 85). Horatio, the master responded to this suggesting her why not reamin her mistress even after he got married. To this, Brown added a text saying, â€Å"True, she was his slave; bones, and sinews had been purchased by his gold, yet she had the heart of a true woman† (Brown 112). Schweninger also points out that Brown inserted Clotel’s consideration of remaining as a mistress a criminal act (Schweninger 28). Another important matter about the novel is the choice of places to mention. Schweninger confirms the accuracy of the accounts in the place where Currer (Clote’s mother) was – Natchez (Schweninger 29). It was a place, according to Buckingham (from Schweninger 29) where gambling and all sorts of vices were rampant, but most especially, it was a place where people were cruel to slaves. Hence, it was also the site where a runaway slave was burned. Another place mentioned that can be noted for accuracy is Richmond, where Clotel’s daughter was sold. Richmond was, according to Marie Tyler-McGraw (in Schweninger 29) the center of slave trade. It was a powerful place, then, primarily because it was the center of slave brokerage. However, there was some mistake in Brown’s choice for the time when Clotel was caught in Richmond. According to Schweninger, Clotel could not have been caught during the time of the Turner uprising in 1831, because she was mentioning names of presidential candidates of 1839 (Schweninger 30). Yet, Schweninger argues that this â€Å"misplacement† is of no importance because what was being pointed out here was the she was accused of being a part of the uprising and that was why she was hiding in disguise (Schweninger 30. ). It clear here that Brown’s intention is to show that slaves, too, are humans, capable of upholding morality, and want to uphold morality. They want to do what is right and to be in a place where they can be free, but the people who hold them captives refuse to let them do what they believe is right. Slaves are capable of love and reason, but all that the people who own them see in them is flesh and blood, not human beings with a soul, heart and mind. They were treated as properties and not as members of the society. I believe it pays to go back to Brown’s own words that the institutionalization and rationalization of slavery stated with the people on top, so to stop slavery, then the â€Å"guilt† should be â€Å"fastened† to the people in the â€Å"higher circle† (Brown iv). When Horatio got married, his wife saw Clotel as a threat so she had her sold. She tried to escape by disguising herself many times. She so got caught by the slave hunters and later committed suicide. This is seen by Articlemyriad. om as the evidence of the idea that the novel does not present optimism or pessimism in whole (Analysis and Themes 1). There were optimistic moments such as those when she was able to escape, yet she still wound up dead by her own will. However, another critique sees the disguises done by Clotel in a different way. Berthold sees every change in costume as a representation of the adversities that Clotel had to face as a female slave (Berthold 19). Guilla argues that male s were connotative for slaves then, so any woman portrayed as a slave would be something new in literature. So, in Berthold’s explanations, he points out that each time Clotel changes costume, there was something to show about the society and something about her personality and slavery (Berthold 19), even making reference to Brown in the 1880 My Southern Home, â€Å"Extravagance in dress, is a great and growing evil with our people† (Brown 232). For instance, most of Clotel’s disguises were as a white male, just to get out of the situation she was in. It is shown here how people are also judged by their clothing, and so goes with the stereotyping, the gender roles and the power in the gender role. So, if we look into the disguises that Clotel had, she first disguised herself as Mr. Johnson, then as an Italian or Spanish. She actually faced more danger when she disguised herself because she was wearing the shoes of someone whose roles she was not familiar with. When she disguised as a Spanish or Italian, the tension must have been great for she had to speak Italian or Spanish if ever any Spanish or Italian spoke to her. It also, then put her life in danger. Yet, the dangerous attempt got her what she wanted. However, in the end, when she committed suicide, Berthold saw this as â€Å"undisguising† herself, one that ascertained freedom for her, for she died as herself, not as anyone else (Berthold 30). Giulla sees the arrangement of the plot as a product of the author’s effort to expose the life of a community of slaves and need â€Å"to structure the fictional text around the evolution of the individual fate (Guilla 639)† and in so doing, explain the shifts from romance to realism in a exhaustively detailed depiction of slavery and the made-up â€Å"re-unions of long lost relatives and lovers (Guilla 639). If we examine the novel, the members of the family were separated because they were sold to different masters and had different fates. They must have yearned to be with each other as a mother wants to be with her daughter and a daughter with her mother, but the cruelty of slavery kept them from being together, even to the point of their death. This brings us back t o the pessimistic ending that Articlemyriad is talking about. In fact, when one is a slave, what optimism can pone really expect, but a life of hard work and subhuman treatment unless, of course one meets an extraordinary master. But even in the case Clotel, she had romance with Horatio, but the man did not consider his affair with her as something valuable, for if he did, he would have agreed to take her to France or England. We now go back to what I said earlier that slaves were seen as flesh and blood but not as humans with soul and mind. Brown’s style in writing the novel presents and extraordinary mixture of fact and fiction. The story is based on a popular lore that the people then knew, yet he gave it a different touch that made the people understand what was going on. Instead of using male slaves as heroes in the story to enable them to engage in physical confrontations and rebellions, Brown used the female slave to show the beauty and morality behind the mulatto and blacks, that though they were slaves, they had good hearts, and they were good people capable of love and ion need of love, especially in need of love. A show of rebellion could have shown what the society would be if slavery continued, but that was something that was expected to happen (although he made a novel end with a civil war). Brown deemed it more important to show, not the violence that will rise if slavery does not stop, but the impact of slavery to the lives of the people who had been being treated as animals for decades and even centuries by people who profess that they are Christians. To this, it makes a reference to the situation when Clotel disguised herself as Spanish or Italian. These people are known for their devotion in Catholicism. Here, I want to point out that the people who profess that they love God are also the ones who have been showing cruelty to the weak. They were actually hypocrites in a time when religion was strictly followed. We can see from the accounts mentioned by Schweninger that Brown had good accounts in geographical locations, but seems to have had some problems with historical accuracy. However, whether he had problems with chronology of events or he intended them to be such to stress certain points (like in the case of Clotel’s capture during the Turner uprising), his work remains to be a living testament of what happened to the blacks and mulattos in the United States. His work remains to be a loud cry during his time, transcending the boundaries of time to portray the evils of slavery. After all that the blacks and mulattos have experienced, they only deserve to have a part in the land once said to be flowing with milk and honey. The land of opportunity belongs, not only to the whites but also the clacks, and their children. Clotel is a literary treasure, not only to the blacks, but also to the world. It is a legacy, which through it history-based narrative has shown the plight of people in a certain place at a certain point of time. It can be considered a dark age of history, but it is also proof that whenever there is something wrong, the right will also shine.

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Long term effects of rape using the Prince of Tides for reference essays For many years troubled adults were ignored or punished for their mental state. Adults who had been raped as children were expected to just "get over it" or "move past it". Society did not connect the fact that the adult had been raped as a child with the problems occurring in adulthood. These adults would display instability in many ways when it came to conducting their lives. In recent history therapists have begun to understand the very real long lasting effect that childhood rape has on the lives of the adult victims. In the story Prince of Tides by Pat Connery the issue of rape effects are explored and explained in the plot. While the book itself is fiction it is a truthful and honest account of the negative impact childhood rape has on the victim when the victim enters and lives in adulthood. This book tells the story of a man who reveals a horrible family tragedy to his sister's therapist(Conroy, 1987). Tom Wingo is one of the adult children in this saga that involves a large cast of characters. This alone is an illustration of the way abuse affects those who are involved. The ripple effect impacts many family members as well as friends of those family members(Conroy, 1987). The book presents the theme that adults are severely and negatively impacted by childhood rape and years later that impact plays an important role in their life choices and mistakes(Conroy, 1987). The story covers a four decade stretch of time thereby illustrating and underscoring the horrors that rape can leave. Other family members are also affected in serious ways and the story discusses the entire family and its dysfunction for the four decades before the story is told. The book dovetails with what the field of mental health has discovered abo...

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William Faulkner essays Many great writers are criticized about the way they write there: novels, poems, essays, etc. Even some of the skilled writers are criticized, William Faulkner was a great writer that wrote several novels but was not criticized that much about the way he wrote. Although Faulkner was very creative in writing his novels and essays he still had various flaws which were sometimes criticized by critics. Before Faulkner won the Nobel Prize in 1950, Faulkner was generally considered by most critics as a minor regionalist and something of a crackpot.(William...). At the time he was really poor and most of his books were out of print. William Faulkner was a young man who never graduated high school, never received a college degree, and was living at the time in the poorest state in the nation. He worked as a screen writer in Hollywood but was known for his drinking binges which affected his health, writings, and relationships with his friends and family. Some of Faulkners best works were: As I Lay Dying, The Sound and the Fury, Light in August, Absalom,Absolom!, and Go Down Moses. Most of Faulkners early writings were romantic, but once he moved to New Orleans he became more popular and began writing several essays and novels. After he finished writing his first novel, Soldiers Pay, he published it and decided to move near Paris, France. Faulkner was mainly influenced be the observations he would take, and experiences of life in the South. Because of the fact that Faulkner grew up in the South being exposed to harsh racism, he witnessed how the blacks were being discriminated against. I believe Faulkner wrote about the South in several of his stories because he was affected greatly by all the racism going on. Growing up around racism, made him write about it in some of his stories, which many people disliked, because he would show racism in his stories. ...

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