Quantitative Real Time Polymerase Chain Reaction (RT-qPCR)

Quantitative Real Time Polymerase Chain Reaction (RT-qPCR) QUANTITATIVE REAL TIME POLYMERASE CHAIN REACTION (RT-qPCR) Primers All primer sequences were designed using the online tool Primer 3-BLAST (NCBI) and the primers were obtained from Sigma Aldrich, Bangalore, India. Relative expression of transforming growth factor beta (TGF- ÃŽ ²), myosin heavy chain beta (ÃŽ ²-MHC), endothelial nitric oxide synthase (eNOS) and glyceraldehydes-3-phosphate dehydrogenase (GAPDH) was studied. Forward and reverse primers for the above genes were used for amplification. Table 5. PCR Primer details RNA isolation All glasswares were rinsed with diethyl-pyrocarbonate (DEPC) treated water to inhibit RNases. Total RNA was isolated using guanidium thiocynate-chloroform-phenol method of Chomczynski and Sacchi (1987). Total RNA isolation kit (BioUltra, Sigma Aldrich,USA) was utilized for this study After cleaning with saline, heart and aorta tissues were homogenized in denaturing solution with freshly added ÃŽ ²-mercaptoethanol. After homogenization 2M sodium acetate solution (pH. 4.0), water saturated phenol and chloroform: isoamyl alcohol (49:1) was added. The mixture was shaked vigorously and allowed to cool on ice for 15 minutes. The mixture was centrifuged at 10,000 Ãâ€" g for 20 minutes at 4 oC. The aqueous phase was transferred in a fresh tube and an equal volume of ice cold isopropanol was added. RNA was precipitated by placing the sample at -20 oC for one hour. Then the mixture was centrifuged at 10,000 Ãâ€" g for 20 minutes at 4 oC. The pellet was washed with 70% ethanol and RNA was stored in DEPC water at -80 oC. RNA quality and quantity was assessed by nano-drop spectrometer. Real time PCR amplification SYBR Green Quantitative RT-qPCR Kit was used in this study and the PCR experiment was carried out in eppendorff realplex mastercycler. 1 µg RNA was reverse transcribed by using Molone murine leukemia virus (M-MuLV) reverse transcriptase as per manufactures instructions. Then the amplification program (94 oC – 45 seconds, annealing – 45 seconds, extension 72 oC- 1 minute) was applied with specific annealing temperature. The annealing temperatures of TGF-ÃŽ ², ÃŽ ²-MHC, eNOS and GAPDH were 58, 52, 55, and 55 oC, respectively. The specificity of the primers was confirmed by resolving the PCR products in 1.5% agarose gel electrophoresis. The relative fold change of expression was calculated by normalized the expression with GAPDH. The RT-qPCR results were quantified using the ‘threshold line’ and the ‘cycle threshold’. The ‘threshold line’ is the point at which the reaction reaches a fluorescent intensity above background. The cycles at which the samples reach this level is called the ‘cycle threshold’ (Ct). The statistical analysis of the RT-qPCR results was calculated by using the à ¢Ã‹â€ Ã¢â‚¬  Ct = (Ct value of gene of interest – Ct value of GAPDH). Relative gene expression was obtained by à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  Ct methods (à ¢Ã‹â€ Ã¢â‚¬  Ct sample – à ¢Ã‹â€ Ã¢â‚¬  Ct of control), with the use of the control group as a calibrator for comparison of all unknown sample gene expression levels. The relative gene expression fold change was derived from 2–à ¢Ã‹â€ Ã¢â‚¬  Ãƒ ¢Ã‹â€ Ã¢â‚¬  Ct (Schmittgen and Livak, 2008). IMMUNOHISTOCHEMICAL LOCALIZATION (IHC) Immunohistochemistry (IHC) IHC was performed as described by Rocha et al., (2009) using Super Sensitive Polymer-HRP Detection System kit, from Biogenex, USA. The Super Sensitive Polymer-HRP Detection System is a atypical detection system using a non-biotin polymeric technology that makes use of two major components: a Poly-HRP reagent and super Enhancerâ„ ¢. As the system is not based on the biotin-avidin system, the problems associated with endogenous biotin are completely eliminated. The detection of antigens in tissues by immunostaining is a two-step process. The first step involves the binding of an antibody to the antigen of interest and the second step involves the detection and visualization of bound antibody by one of a variety of enzyme chromogenic systems. The choice of detection system will dramatically impact the sensitivity, utility and ease-of-use of the method. Procedure Paraffin-embedded tissue was cut to obtain sections of about 4  µm thickness. The mounted paraffin-embedded slices are deparaffinized in xylene and rehydrated using an ethanol/H2O gradient. Heat mediated antigen retrieval step was carried out for 10 min and then the slides were allowed to cool to room temperature for another 20 min. This was followed by peroxidase block treatment (to block endogenous peroxidase enzyme activity) for 10-15 min and then power block treatment (to block non-specific binding of antibodies to highly charged sites) for another 15 min. The sections were incubated with the concerned diluted primary antibody solution (for 2 h (1:200)) followed by treatment with the super enhancer solution (for 30 min) and super sensitive Poly-HRP solution (for 30 mins). After colour development with DAB and counterstaining with haematoxylin, the sections were observed under the microscope and photographs were taken. TRANSMISSION ELECTRON MICROSCOPIC STUDY The ultrastructure of the heart specimen was examined by Transmission Electron Microscopy (TEM) according to the method of Lang (1987), by the technique of thin sectioning. Reagents Glutaraldehyde solution: 3% Osmium tetroxide: 2% osmium tetroxide in 10 mM sodium phosphate buffer, pH -7.4 Ethanol: 75%, 95% and 100% Uranyl acetate: 1% Lead citrate: 3% Sodium phosphate buffer: 0.1 M, pH 7.4 Procedure Immediately after the sacrifice, the heart tissues were dissected and fixed with a solution of 3% glutaraldehyde for 2 hours at room temperature and washed thrice with phosphate buffer to remove glutaraldehyde. Post-fixation was done by a solution containing 2% osmium tetroxide in 10mM sodium phosphate buffer and left overnight. Then, the osmium tetroxide solution was removed and replaced with 75% ethanol. This reduces the remaining osmium tetroxide to osmium dioxide, which forms a precipitate in the alcohol. After 10 minutes, the alcohol was replaced with a few ml of 75% ethanol. After 30 minutes, the alcohol was replaced with 95% ethanol and left for 30 minutes. This solution was replaced with 100% ethanol and washed thrice and then dried in acetone. After dehydration, the tissues were equilibrated for 30 minutes in 1:1 mixture of epoxy propane and the embedding medium, epon 812 (also called epikote resin-812). A mixture of the resin and two hardening agents, dodecyl succinic anhydride and methyl anhydride were used. A diamine catalyst generally N-benzyl-N-diethylamine was added just before use. The 1:1 mixture was poured off and replaced with full strength resin. This step was repeated several times to ensure full infiltration of the embedding medium. The tissue was then transferred to a beam capsule with a wooden stick and the capsule was filled with fresh resin mixture. The wooden stick was used to tease the specimen down to the center of the bottom of the capsule. Next, the block holder was placed with the specimen in hot air oven at 60 °C for 48 hours to polymerize the resin completely. Once the blocks are hardened, they are ready for sectioning. The ends of the specimen blocks were trimmed using glass knives and ultra thi n sections were cut using an LKBUM4 ultramicrotome. The sections were picked upon carbon grids and post-stained with combined uranyl and lead stain and rinsed with distilled water and dried. After drying, the grids were examined under a Philips EM201C transmission electron microscope (Philips, Eindhoven, Netherlands). WESTERN BLOT ANALYSIS Western blotting was performed to analyze the expression pattern of eNOS in the aorta and reperfused hearts according to method of Laemmli (1970). Principle Following the protein estimation, the samples were separated using SDS-PAGE gel electrophoresis and the separated molecules are blotted onto a polyvinylidene fluoride (PVDF) membrane. After blocking, the primary antibody was added and allowed to bind to the protein followed by washing (which removes non specifically bound antibody); then an enzyme-labeled secondary antibody was added, to detect the primary antibody. The location of the secondary antibody was determined by adding an appropriate substrate for the enzyme conjugated to the secondary antibody. Reagents Acrylamide stock: 30% acrylamide, 0.8% N,N†²-methylene bisacrylamide Separating gel buffer: 1.5 M Tris, pH 8.8 Sample buffer: 0.5 M Tris, pH 6.8 Sodium dodecylsulfate (SDS): 10% Ammonium per sulfate (APS): (10%) N,N,N,N-tetramethylethylenediamine (TEMED) Separating gel overlaying solution: Water-saturated isobutanol Sample Buffer: Tris (0.5M, pH 6.8)-2.5 mL SDS (10%)-4.0 mL Glycerol (100%)-2.0 mL ÃŽ ²-Mercaptoethanol-0.8 mL (or 1 M DDT-0.5 mL) Bromophenol Blue (0.1%)-300  µL Distilled water (400  µl) to 10.0 mL Running gel buffer Tris-6.05 g Glycine: 28.80 g 10% SDS: 10.0 mL or (1.0 g) Distilled water to 1000 mL Staining solution Coomassie brilliant blue R250- 300 g Methanol-80 mL Acetic acid-20 mL Distilled water-100 mL Destainning solution Acetic acid-100 mL Methanol-300 mL Distilled water: 1000 mL Procedure The aortic tissues were homogenized in an ice-cold radio immuno precipitation buffer (RIPA) (1% Triton, 0.1% SDS, 0.5% deoxycholate, 1 mM/L EDTA, 20 mM/L Tris (pH 7.4), 150 mM/L NaCl, 10 mM/L NaF, and 0.1 mM/L phenylmethylsulfonyl fluoride (PMSF)). The homogenate was centrifuged at 10,000 Ãâ€" g for 20 min at 4 °C to remove debris and the supernatant was used to determine the protein concentration of the lysates using the BCA protein assay kit (Merck, India). Transfer of proteins to membrane Samples containing 50 ÃŽ ¼g of total cellular proteins were loaded and separated using 10% SDS polyacrylamide gel electrophoresis. Following electrophoresis, the proteins were transferred from the gel to a membrane by using semi-dry blotting system (AA Hoefer, SEMIDRY BLOTER, USA). Before assembling the transfer system, soaked PVDF membrane in methanol for 10 minutes and blotting papers in cold transfer buffer. Prepared sandwich, blotting paper, membrane, gel and blotting paper, were placed in the transfer apparatus and few drops of transfer buffer was added and subjected to an electric current 20 V for 1 h under cold condition. After the transfer, the sandwich was removed from the transfer system. Membrane was stained with 0.5% ponceau in 1% acetic acid to confirm equal loading and then washed with distilled water. The PVDF membrane were blocked with 5% blocking solution (containing 5% BSA in 0.5 M Tris-buffered saline, pH 7.5) for 2 h to reduce the non-specific protein binding sites and then incubated with primary antibody (anti-eNOS), in blocking solution with gentle shaking overnight at 4 °C. After this, the membranes were washed with TBST (Tris-buffered saline and 0.05% Tween-20 (TBST)) thrice for 10 minutes interval and then incubated with respective secondary antibody anti-mouse IgG (Sigma-Aldrich, USA) conjugated to horseradish peroxidase. Then the membranes were washed with TBST thrice for 10 minutes interval. The reaction was developed with a DAB detection system (Merck, India). Bands were scanned using a scanner and quantitated by Image J, a public domain Java image processing software, Wayne Rasband, NIH, Bethesda, MD, USA. H9c2 cardiomyoblast cell culture Rat embryonic cardiomyoblast derived H9c2 cells was obtained from National Centre for Cell Science (NCCS), Pune, India. Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum and a combination of penicillin-streptomycin (1%) in a humidified 5% CO2atmosphere at 37 °C. The assay was performed by seeding H9c2 cells in the concentration of 1Ãâ€"104 cells/well in 96-well plate. In vitro oxidative stress and mitochondrial transmembrane potential study In order to evaluate the cytotoxic effect, viability was checked with MTT assay on D-carvone (25–100  µM) treated H9c2 cells. For assessment of protective potential of D-carvone against oxidative stress, different concentrations of D-carvone (0.1, 1 and 10  µM) were incubated with H9c2 cells for 2 h, and then co-incubated with 500  µM/L H2O2 for further 18 h (Jia et al., 2012; Zhang et al., 2011). For viability analysis, MTT solution (5 mg/mL) was added to each well, and incubated for 4 h at 37 °C. After incubation, optical density (OD) was measured on a microplate reader at 570nm. With the 10  µM dose of D-carvone, the level of intracellular ROS formation was quantified with fluorimetry using redox-sensitive fluorescent probe 2, 7-dichlorodihydrofluorescin diacetate (DCFH-DA). Further, to examine mitochondrial membrane permeability transition (MPT), H9c2 cells were incubated with 5 mg/mL Rhodamine 123 (Rh123) at 37 °C for 30 minutes (Park et al., 2003). The images were acquired using the Olympus IX71 inverted à ¯Ã‚ ¬Ã¢â‚¬Å¡uorescence microscope. Ischemia/reperfusion (I/R) protocol D-carvone was dissolved in 1% DMSO (vehicle) and administered orally to rats using an intragastric tube daily for 7 days. The rats were randomly divided into four groups of six rats per group: (i) control group pre-treated with vehicle alone for 7 days (isolated rat hearts subjected to continuous perfusion). Isolated rat hearts obtained from the following three groups were perfused with a modified Krebs buffer solution for 10 minutes to stabilize the cardiac functions and then subjected to 30 minutes of global ischemia, followed by 60 min of reperfusion: (ii) I/R hearts pre-treated with vehicle alone for 7 days (Control (I/R)); (iii) I/R hearts pre-treated with D-carvone (I/R + D-C 10 mg/kg body weight); (iv) I/R hearts pre-treated with D-carvone (I/R + D-C 20 mg/kg body weight). Langendorff isolated heart preparation The animals were anaesthetized with an intramuscular injection of ketamine (75 mg/kg body weight). After thoractomy, the hearts were rapidly excised and placed in cooled (4 °C) Krebs Henseleit bicarbonate solution [composition (in mM): 118 sodium chloride (NaCl), 4.7 potassium chloride (KCl), 1.2 magnesium sulphate (MgSO4), 1.2 potassium dihydrogen orthophosphate (KH2PO4), 2.3 calcium chloride (CaCl2), 25.0 sodium bicarbonate (NaHCO3), 11.0 glucose]. composition (in mM): 118 sodium chloride (NaCl), 4.7 potassium chloride (KCl), 1.2 magnesium sulphate (MgSO4), 1.2 potassium dihydrogen orthophosphate (KH2PO4), 2.3 calcium chloride (CaCl2), 25.0 sodium bicarbonate (NaHCO3), 11.0 glucose. The heart was then attached to the cannula through aorta and retrogradely perfused with the Krebs solution maintained at 37 °C and continuously gassed with a mixture of 95% O2 5% CO2. Perfusion pressure was kept constant at 80 mmHg. The ischemia and reperfusion protocol was followed as described previously (Khan et al., 2006; Senthamizhselvan et al., 2014). An elastic water-filled balloon was introduced into the left ventricle through a left atrial incision and connected to a Pressure Transducer (AD Instruments) linked with a PowerLab data acquisition unit (AD Instruments). The balloon volume was adjusted to achieve a stable left ventricular end-diastolic pressure (LVEDP) of 5-10 mmHg. The percentage rate-pressure product [RPP = (LVSP-LVEDP) Ãâ€"HR] and percentage coronary flow was assessed as described previously (Esterhuyse et al., 2005; Ferrera et al., 2009; Swaminathan et al., 2010). Coronary effluent was collected for the estimation of LDH activity. Macroscopic enzyme mapping of infarcted myocardium (Triphenyl Tetrazolium Chloride test) TTC (triphenyl tetrazolium chloride test) test used for a section of the heart tissue. Lie et al. (1975) method was used for the triphenyl tetrazolium chloride test (TTC) analysis acclimated for the macroscopic enzyme mapping appraisal of the infarcted myocardium was completed. A freshly prepared solution of 1% TTC in phosphate buffer was prewarmed at 37-40 °C for 30 minutes in a darkened glass. To remove the excess blood, the heart tissues were washed rapidly in cold water without macerating the tissue. After removing epicardial fat, the left ventricle was taken separately. To obtain slices not more than 0.1-0.2 mm in thickness, the heart was transversely cut across the left ventricles. The heart tissue slices were kept in the covered, darkened glass dish containing prewarmed solution of TTC and the dish was kept in an incubator and heated to 37-40 °C for 45 minutes. The heart slices were turned over thrice and made certain that it remains fully immersed in the TTC solution. At the end of the incubation period, kept the heart slice in fixing solution to fix the tissue. Colour photographs of slices were obtained by a camera with macro lens. The expected reaction of the TTC test was as follows: normal myocardium (LDH enzyme active) turned to bright red, infarcted myocardium (LDH enzyme deficient) turned to uncolored white.

The R-word and Racist Native American Sports Team Logos Essay

Racial epithets have long existed and plagued our society, Native Americans throughout the country consider the R-word a racial, derogatory slur along the same lines of other hurtful, slanderous, and offensive ethnic insults including the N-word among African-Americans, the K-word for the Jewish and the W-word amongst Latinos. Above all, the portrayal of stereotypical Indian images is common in American popular culture (i.e. Jeep Cherokee, Land O’Lakes butter). Moreover, the use of Indian logos or mascots at both the professional and high school level in sports has become increasingly controversial. Thus, the removal of Native American mascots from sports teams is necessary to fight the injustice of the negative connotations and stereotypes that are typical in the depiction of Indians. Our society must become aware of how very racist the word â€Å"redskin† is and how very derogatory the portrayal of the Native American is in so many commercial and sporting events. Interestingly, Merriam-Webster’s definition defines â€Å"Redskin† as a very offensive slang used as a disparaging term for a Native American and should be avoided. The fact that many Americans are not aware of the definition of the term â€Å"redskin† or are blind to see into believing that this term means strong, brave, and courageous gives them a false sense of understanding to the true testament of the word â€Å"redskin† that is heavily misunderstood and overlooked in today’s society. First, by considering the term â€Å"Redskin† has for centuries been used to belittle and humiliate an entire people. The meaning originated in colonial times when traders and local government paid for skins. There was a certain price paid for various animal skins. On that list was the term â€Å"Red-skin,† which referred to bloody scalps of American Indians resulting from a Native American crossing the path of a bounty hunter. Most of the affected tribes were Penobscots, Passamaquoddy, Wampanoag, Mashpee Wampanoag and others along the New England coastal line. The reason they were paid for these scalps, the colonists were working to remove the American Indian presence and take over their land. Furthermore, the original name was a European one used to describe Algonquins who painted their face with bright red ocher and bloodroot, consequently making their  face red with war paint. In addition, red is the most common color used by Native Americans in painting their skin. According to Dress Clothing of the Plains Indians by Ronal P. Koch, â€Å"Red is generally accepted as being one of the colors most easily available to and most used by Indians for decorative and ceremonial purposes.† In recent developments, the Non-Disparagement of Native American Persons or Peoples in Trademark Registration Act of 2013 (H.R. 1278) introduced by U.S. Congressman Eni Faleomavaega of the Territory of American Samoa states that this bill would require cancellation of existing trademark registrations for trademarks using the term â€Å"redskin† in reference to Native Americans. It would also deny registration for new trademarks so using the term â€Å"redskin† would be deemed improper, the bill has begun to pick up steam and has garnered nation wide support through the backings of Native Americans and Non-Native American organizations in advocating an end to the use of the term â€Å"redskin† which constitutes a racial slur and is disparaging, derogatory, demeaning, and offensive to Native Americans. According to the United States House of Representative’s website, documented in a letter to Members of Congress, the National Congress of American Indians (NC AI) which is the oldest, largest and most representative American Indian and Alaska Native organization serving tribal governments and communities recently stated: This legislation will accomplish what Native American people, nations, and organizations have tried to do in the courts for almost twenty years – end the racist epithet that has served as the [name] of Washington’s pro football franchise for far too long. The Tulsa Indian Coalition Against Racism (TICAR) asserts that the â€Å"R-word† is â€Å"hurtful and injurious to our youth, as well as the entire Native American population.† Accordingly, the Native American Finance Officers Association (NAFOA) affirms: The term has never been acceptable in the Native community and causes harm to the collective self-esteem and status of American Indians in the larger society. . . What should be viewed as a national embarrassment has somehow turned into a  celebrated namesake for a national sport? Further, the American Indian Movement West (AIM-WEST) sustains that: Our organization supports the goal of ridding the sports world of the disparaging name of the Washington pro football franchise. There is no question that this is a racist term that causes harm and injury, whether or not it is intended to do so, and must not be tolerated in decent society. As well as, the United South and Eastern Tribes, Inc. (USET) expresses: Overcoming the social challenges resulting from industry branding and media exposure has taken generations for other groups. Native communities are on a similar journey. In our work to protect and promote our sovereignty rights at all levels, existing stereotypes, bigotry, and racist views about our people often get in the way of progress. This legislation will assist Tribes in promoting an understanding of American Indian culture, positive images of Indian Country, the effects of historic trauma, and the modern-day successes and challenges Tribes face as we seek to improve the standard of living within our communities. In addition to the above organizations, there are 50 other organizations that have either pledged their support for this bill or rejected the use of the term ‘Redskin,’ among them are the Cherokee Nation of Oklahoma, the Comanche Nation of Oklahoma, the Oneida Indian Tribe of Wisconsin, the Seminole Nation of Oklahoma, the National Indian Youth Council, the National Indian Child Welfare Association, the American Indian Higher Education Consortium, the National Indian Education Association, the National American Indian House Council as well as a long laundry list of other notable organizations. Second, the stereotypical Indian images in American pop culture known as â€Å"Tribalism,† as Ruth Hopkins, a columnist for the Indian Country Today Media Network concedes â€Å"as a mainstream trend largely based on false, stereotypical notions of who indigenous people are has become a pop culture phenomenon.† Among those are celebutantes, pop princesses and hipster  wannabes have been wearing obtrusive, exaggerated war bonnets and headdresses, wearing â€Å"war paint,† and playing dress up in Native American â€Å"inspired† costumes in record numbers. She goes on to illustrate that the perpetuation of stereotypical images of Native peoples is unacceptable and discriminatory for a plethora of reasons. (Hopkins) Clearly, Non-natives who wear American Indian costumes are pretending to be someone of another race. Those who play â€Å"dress up† by wearing an American Indian costume, headdress or war bonnet are not only failing to acknowledge the existence of over 500 recognized Native nations, each separate and distinct from one another, they are making light of centuries of suffering, oppression and massacre endured by the indigenous people of this country. Enforcing racial stereotypes of Native peoples as savages in characterless feathers and fringe also perpetuates the myth that American Indians are not active members of modern society and casts them aside to make them feel further disrespected and unworthy as a depleted and lost society. (Hopkins) Actually, not all American Indian tribes include war bonnets or headdresses as part of their traditional insignia. Of those who do, headdresses and war bonnets were worn by men, and have nothing to do with fashion or the sexual objectification of women. Hopkins expresses that â€Å"each eagle feather contained in a war bonnet is individually earned, often bestowed upon the owner through ceremony, and represents a significant event or acknowledged act of bravery, leadership, or self-sacrifice.† Much less, powerful, respected American Indian men with a history of valor who are leaders in their Tribal community specifically wear war bonnets. In other words, the only people who should be wearing war bonnets are chiefs or well respected warriors, such as Tatanka Iyotanka or Chief Sitting Bull of the Hunkpapa Lakota not these so-called reality stars such as Chloe Kardashian, or the pop star Ke$ha. It’s sad and pathetic that such an inconsiderate display would be similar to th e wearing of a war bonnet by someone who hasn’t earned it. (Hopkins) Because many people have such a limited knowledge of Indians, Native Americans are arguably, among the most misunderstood ethnic groups in the United States. Native Americans are also among the most isolated groups. What people know is limited by their sources of information and,  unfortunately, much of the information about Indians is derived from popular culture. Stereotyping is a poor substitute for getting to know individuals at a more intimate, meaningful level. By relying on stereotypes to describe Native Americans, whites come to believe that Indians are drunks, get free money from the government, and are made wealthy from casino revenue. Or they may believe that Indians are at one with nature, deeply religious, and wise in the ways of spirituality. Indeed, American mainstream media have always tended to distort Native American images. In a research conducted by Liu & Zhang on the representation of Native Americans in pop culture, â€Å"the film Dances with Wolves; the radio and TV Western, The Lone Ranger; and the novel, by Fenimore Cooper, The Last of the Mohicans, are just a handful of TV shows ad movies that present negative or romanticized images of American Indians, either nasty or cruel, or subservient and short, but all disappearing.† For instance, the fabricated Indian images on TV and in the Hollywood films influence the identity formation of individual Native Americans. Consequently, Hollywood and TV have created simulated Indians and have played and replayed these images so many times that the Indian viewers take them as real. These romantic and stoic characters hardly speak in the films; nor do they get heard. Especially, Liu & Zhang point out the discrepancies that in Hollywood films and TV plays, Indians ar e paid to die, to fall off the horse, to confirm the â€Å"Vanishing Noble Savage† stereotype, so endings are important. Further propagating that these stereotypical images can be seen in the â€Å"westerns† movies and even in some cartoons such as Peter Pan. Moreover, other stereotypical images showed them with painted faces smoking peace pipes, dancing around a totem pole (at times with a captive tied to it), sending up smoke signals, wearing feathered head pieces, scalping the heads of their enemies and constantly chanting the word â€Å"um† promotes a damaging misconception and negative inferences towards Native Americans. With regards to discrimination, when the highly popular Twilight series received the Hollywood treatment, Taylor Lautner played the Native American character Jacob Black and his casting became steeped in controversy. As Dow points out that â€Å"Lautner’s presence seemed out of sync with Hollywood’s recent pro-Indian stance. Lautner claimed to have discovered his Indian  ancestry after being cast. Actions like this show film producers’ hesitance to hire an actor in spite of the character’s ethnicity. Rick Mora, an actor who resides in California, who plays a Native American in T wilight disagrees with the casting of Taylor: â€Å"There is plenty of Native talent in town (Hollywood) to play that role.† Furthermore, she yields that the movie could be â€Å"applauded for representing Natives as more than simply a dying race, instead appearing onscreen as people with their own unique personalities.† For some younger viewers this may be their first contact with Native American culture, so acknowledging Indians as Americans on screen was an achievement on the part of Hollywood. In addition, the summer release of X-men Origins: Wolverine in 2009 and the highly popular character Silver Fox made her first onscreen appearance in the movie series. In the original comics, Silver Fox is described as a Native Canadian Black Foot. The character is to be played by Caucasian actress Lynn Collins, and the decision to cast a white actress has upset many fans of the comics. Hollywood producers have also decided to change her name to the more American-sounding Kayla Silverfox. Clearly, not only does Hollywood still find it difficult to include a Native American in a blockbuster, but also they e ven refuse to leave the traditional ethnic names intact. (Dow) Whereas names, images, and mascots that symbolize Native Americans are used extensively in the United States, particularly in sports and advertising. In sports there are the Washington Redskins football team, the Atlanta Braves and Cleveland Indians baseball teams, and the Chicago Blackhawks hockey team. Fans of the Atlanta Braves use the â€Å"tomahawk chop† accompanied by a chant to intimidate visiting teams, while the Cleveland Indians use the mascot Chief Wahoo and the University of Illinois uses the mascot Chief Illiniwek. As a result, Native Americans across the country have been protesting the use of their symbols and heritage in sports arenas for over a decade. Most particular in the realm of professional sports, these protests have not generated significant changes in attitudes and practices. As an illustration, Hatfield designates that logos used by the Washington football team and the Cleveland and Atlanta baseball teams are offensive for many reasons, as are the logos formerly used by Dartmouth College and the University of Illinois. (They are no longer used because the NCAA banned teams with racist names and ma scots from post-season play.) He implies that  these logos appropriate the identities of Native Americans, many of whose languages and cultures have been destroyed by Euro-Americans. They take sacred religious symbols from Native American cultures – eagle feathers, face paint, and peace pipes – belittle them, and exploit them for the commercial and entertainment purposes of Americans. And they perpetuate outdated, demeaning stereotypes of Native Americans that make it difficult for Native Americans to represent themselves as part of contemporary American society.  Be that as it may, these logos reduce Native Americans to savages, to defeated enemies who have been â€Å"erased† from today’s world. Indian mascots objectify and commercialize Native Americans and their cultures. Cigar store Indians were used as advertisements to sell tobacco. Urban Outfitters used Navajo patterns to sell clothes, at least until lawyers representing the Navajo Nation filed suit against them and won an i njunction forcing them to stop. (Hatfield) Other nicknames of professional and college teams, such as Indians, Braves, Chiefs, and Seminoles may not in themselves be offensive. However, the portrayal of these words is often very demeaning. For example, the 1995 World Series, the Cleveland Indians and the Atlanta Braves, with Chief Wahoo as the mascot for the Cleveland team and the â€Å"tomahawk chop† exemplified by fans of the Atlanta team, portrayed Native Americans in an extremely degrading manner. Suzan Shown Harjo, Director of the Morning Star Institute, says that this portrayal of Native Americans is â€Å"racist, derogatory, demeaning, pejorative, offensive and ignorant at best.† On the other hand, Dr. Cornel Pewewardy, a visiting scholar in the Department of Education at Cameron University, has written extensively about the struggle of unlearning ‘Indian Stereotypes’ for both Native Americans and non-Native Americans as learned from the demeaning public portrayal of the American Indian through mascots, the movie, Pocahontas, and the â€Å"tomahawk chop.† Being that there are 62 high schools that use the name Redskins, the term has vanished from the collegiate landscape. Accordi ng to Capital News Service, â€Å"the last two colleges that used Redskins changed the name in the late 1990s. Miami University of Ohio changed from the Redskins to RedHawks in 1997 and the Southern Nazarene Crimson Storm dropped the name in 1999. If the two universities had not changed their name by 2006, they would have been unable to play in the postseason under a NCAA policy adopted in 2005 that bans the use of Native American mascots by sports teams during its tournaments.† The postseason ban convinced colleges with mascots like Braves, Indians and Savages to become the Red Wolves, War Hawks, Mustangs or Savage Storm. In view of the fact, the CNS denotes that the policy made an exception for teams that have the consent of local Native American tribes like the Florida State University Seminoles. At the high school level, there is no single national sports organization like the NCAA to pressure schools to abandon Native American mascots. But officials in a growing number of states are taking similar steps as the NCAA to force schools to change. Wisconsin passed in 2010 the nation’s first state law banning public schools from using Native American names, mascots and logos. It left exceptions for schools that had the approval of local Native American tribes. In 2012, the Oregon State Board of Education issued a ruling banning all Native American team names, mascots and logos. Affected sc hools must comply by 2017 or risk losing state funding. Alternatively, according to Munson, â€Å"Indian† logos and nicknames create, support and maintain stereotypes of a race of people. She asserts that when one or many of society’s institutions support such cultural abuse, it constitutes institutional racism. Further, the logos, along with other societal abuses and stereotypes separate, marginalize, confuse, intimidate and harm Native American children. They create barriers to their learning throughout their school experience. Additionally, the logos teach non-Native American children that it’s all right to participate in culturally abusive behavior. Children spend a great deal of their time in school, and schools have a significant impact on their emotional, spiritual, physical and intellectual development. As long as such logos remain, both Native American and non-Native American children are learning to tolerate racism in our school. Understanding the history of Native Americans is important to understanding why this is such a controversial topic. The Native American community for 50 years has worked to banish images and names like Chief Wahoo, Washington Redskins, Kansas City Chiefs and the Atlanta Braves. It is important to remind people of the cognizant use of the symbols’ resemblance to other historic, racist images of the past. She adds that Native Americans struggled to survive in harsh situations. The support of these mascots only  brings back memories of their ancestors and the suffering and pain they went through for their children and grandchildren. The debate is about more than sports teams and what they call themselves; it is about how Americans treat one another. It is about the respect that different ethnic groups have for those different than themselves in terms of history, physical characteristics, values, and most importantly, emotions. (Munson) In essence, I have came to the conclusion that the Washington Redskins were originally known as the Newark Tornadoes and then the Boston Braves. Most accounts can agree that team owner George Preston Marshall changed the franchise name from the Boston Braves to the Boston Redskins in 1933 to recognize then coach, William â€Å"Lone Star† Dietz. Dietz, who claimed half-German, half-Sioux background, embraced what he perceived to be a Native American heritage. So, since many Native Americans are outraged about the symbolization of Native Americans in sports and advertising, and since society would not tolerate equivalent symbols of other minorities, it is clear that Native Americans are discriminated against, regardless of how others may feel about the matter–and that their civil rights are violated by such racial discrimination. These are important reasons for eradicating the use of Native American names in sports, advertising, and elsewhere. Consequently, Native American organizations such as the National Congress of American Indians (NCAI) are making a strong push through legal action in a bid to force the Washington Redskins to change their name. Most notable of these cases are Pro Football vs. Harjo and Blackhorse v. Pro-Football, Inc. that have made strong efforts in the fight against the discrimination of Native Americans. Works Cited Hatfield, Dolph L. â€Å"The Stereotyping of Native Americans.† The Humanist Sept. 2000: 43. Opposing Viewpoints In Context. Web. 15 July 2013. Washington, d.c.—members of congress urge snyder and the national football league to change the washington team’s name. (2013, May 28). Retrieved from http://www.house.gov/apps/list/press/as00_faleomavaega/eniredskins.html Miller, Jackson B. â€Å"Indians, Braves, And Redskins: A Performative Struggle For Control Of An Image.† Quarterly Journal Of Speech 85.2 (1999): 188. Academic Search Premier. Web. 15 July 2013. Soong, Kelyn. â€Å"The Other Redskins.† . Capital News Service. Web. 15 Jul 2013. . Koch, Ronald P. Dress Clothing of the Plains Indians. University of Oklahoma Press, 1977. Examination of the design and construction of Plains Indian formal †¦ www.minnesotahumanities.org/Teachers/3-04plains.htm Hopkins, Ruth. â€Å"Indian Country Today Media Network.†Tribalism as Pop Culture Phenomenon and the Perpetuation of Offensive American Indian Stereotypes. N.p., 19 Aug 2011. Web. 14 Jul 2013. . Liu, Kedong, and Hui Zhang. Self- and Counter-Representations of Native Americans: Stereotypical Images of and New Images by Native Americans in Popular Media. Harbin Institute of Technology, China, n.d. Web. 15 Jul 2013. . Dow, Madeline. â€Å"Race, Gender, and Mass Media Blog.†Native American Portrayal in Cinema. N.p., 06 Nov 2012. Web. 14 Jul. 2013. . Munson, Barabara. Common Themes and Questions About the Use of â€Å"Indian† Logos. N.p., n.d. Web. 14 Jul 2013. .

PSY Assignment

I think that more than half of the student population at accredited u enlistees have tallest tried a drug that would enhance their focus for the sake of doing well In one of more classes. Step 2: (Hypothesis): The hypothesis Is that more than half of the student population at credited universities have used a performance or cognitive enhancer to do well In one of more classes.Step 3 (Predictions): Possible outcomes for this experiment is that 1) None of the stud .NET are aging performance or cognitive enhancing drugs 2) None of the students are admit Eng to taking performance or cognitive enhancing drugs 3) All or some students will admit to taking g performance or cognitive enhancing Step 4 (Research Method): For this experiment it would be best to do an Survey.A sure very would allow the participants to remain anonymous if they wish to do so and this mix HTH also incline them to be more honest. Step 5 (Subject population) : Age: 1823, Gender: Both male and female, Education: underg raduate and/ or In an undergraduate program, Location: University of Arizona, Arizona State university and Northern State University.Today a research method benefits me because It helps me understand how to proper lay collect and record data to find the results of any question that I want the answer to. With the psychology research method It Is easier to effectively test subjects while upholding all the ethical guldens set by the American Psychological Escalation (PAP). Since all the steps are easy to fool low as of today feel that I can successfully pick a topic that I want to research and find an NAS were to it.

The Influx Of Immigrants During The United States Essay

The influx of immigrants in the United States has been a source of much controversy since the 1790s. Throughout U.S. history there have always been immigration waves shaping the respective time period. For example, the 1880s were characterized by an increase of eastern and southern Europeans, while post-1965 has seen an increased presence of immigrants primarily from Latin America and Asia (Barone 12). Each wave of immigrants adds to the diversity of the U.S. population by bringing their own languages, religions, customs, culture, etc. Despite the open-door policy that once prevailed in America, each of these groups faces prejudice from some Americans who feel threaten by their presence. That should not be the case for some many newcomers having to readjust to a better life in the United States. Americans critics, who themselves are descendants of immigrants, worry that immigration changes the size and composition of the U.S. population. Furthermore, American’s ambivalence tow ards this group has made the treatment of illegal immigrants a social injustice issue that the United States has fail to acknowledge with concrete solutions. Today an estimated of about 11.9 million immigrant make up the population living in the United States. As a result, a large portion of Americans have negatively associated undocumented immigration to opportunity loss for American citizens; however, the undocumented community has positively contributed to the shaping of American history,Show MoreRelatedGlobal Influx Of Immigrants During The United States1074 Words   |  5 PagesGlobal Influx of Immigrants In Texas June 5, 2015, A Salvadoran, Mauricio Hernandez, was sentenced to 50 years in prison and faces deportation after his term for raping his own baby (Immigration Issues 2015, Par. 3). 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